Validation of computer-assisted sperm-motility analysis in the amphibian Silurana tropicalis

2015 ◽  
Vol 27 (7) ◽  
pp. 1049 ◽  
Author(s):  
Severine Larroze ◽  
Daniel B. Pickford ◽  
William V. Holt
Keyword(s):  
Sperm Motility ◽  
Treatment Effects ◽  
Computer Assisted ◽  
Chi Square ◽  
Sperm Activation ◽  
Coefficients Of Variation ◽  
Silurana Tropicalis ◽  
Tracking Time ◽  
Multivariate Pattern ◽  
Objectively Measured

We have developed and validated a computer-assisted sperm-motility assessment (CASA) method for use with the emerging amphibian model Silurana tropicalis. The testicular sperm-activation method was validated by analysing activation replicate coefficients of variation, effects of tracking time settings on velocity distributions and the relative partitioning of differentially motile sperm subpopulations between matched right and left testes. Two major sperm subpopulations were identified using multivariate pattern analysis and their relative frequencies were consistent between samples from matched right and left testes and from randomly drawn subsets of six frogs sampled from the total set of 16 frogs. The power of this approach for detecting treatment effects targeting the hypothalamic–pituitary–­gonadal axis was investigated by injecting a group of frogs with 100 IU human chorionic gonadotrophin (hCG) 2 h before sampling and comparing their sperm-subpopulation frequencies with non-injected controls. While parametric analysis across sperm samples failed to detect treatment effects, subpopulation analysis showed that hCG significantly increased the proportion of progressive and non-sinuous spermatozoa compared with controls (Chi square = 6.40, DF = 1, P = 0.011). This demonstrated the potential value of analysing objectively measured sperm behaviour as an endpoint.

231 SPERM SUBPOPULATIONS IN THE KOALA (PHASCOLARCTOS CINEREUS) EJACULATE

2009 ◽  
Vol 21 (1) ◽  
pp. 213
Author(s):  
N. Satake ◽  
S. D. Johnston ◽  
W. V. Holt
Keyword(s):  
Sperm Motility ◽  
Sperm Head ◽  
Computer Assisted ◽  
Multivariate Pattern Analysis ◽  
Phascolarctos Cinereus ◽  
Non Linear ◽  
Group 3 ◽  
Group 2 ◽  
Head Morphology ◽  
Multivariate Pattern

Koala semen contains a heterogeneous mixture of sperm morphotypes, mainly attributable to extreme degree of shape variability displayed by the hooked sperm head. By analogy with other species, we anticipate that the morphotypes may exhibit correspondingly different sperm-motility behaviors, largely caused by the differences in hydrodynamic interactions with the suspending media. This trend has been shown in human spermatozoa where motility behavior was demonstrably correlated with the sperm head morphology (Overstreet et al. 1981). In this study, we have investigated the heterogeneity of koala sperm motility profiles in semen in an effort to determine whether distinct sperm subpopulations within ejaculates are recognizable by the use of computer-assisted sperm motility analysis. Ejaculates from 5 males were collected by electroejaculation, then diluted and transported in Tris-citrate-glucose (TCG) diluent. Spermatozoa were washed through a 35–60% Percoll gradient to separate seminal plasma and the majority of the prostatic bodies from spermatozoa. Spermatozoa from the washed pellet were then diluted in TCG at 35°C, incubated for 10 min, and video recorded using a negative phase ×10 objective. Sperm motion parameters were then analyzed using the Hobson sperm tracker (Hobson Vision Systems, UK: Holt et al. 1996 J. Androl. 17, 587–596). Multivariate pattern analysis (PATN; CSIRO Australia; Abaigar 1999 Biol. Reprod. 60, 32–41) was used to distinguish 3 sperm subgroups, consistently shown in each ejaculate, within the data (1936 tracks × 6 kinetic parameters; VCL, VAP, MAD, BCF, ALH, LIN). After group allocation by PATN, all parameters showed significant differences between each of the groups (P < 0.0001). Group 1, approximately 25% of the sperm tracks, showed profiles of spermatozoa with fast, non-linear motility (VCL 106.88 ± 28.15; BCF 3.23 ± 3.81; LIN 14.08 ± 10.20). Group 2, approximately 27% of sperm tracks, showed profiles of fast, linear motility (VCL 63.92 ± 13.50; BCF 7.90 ± 3.42; LIN 28.10 ± 12.15). Group 3, 48% of sperm tracks, showed profiles of slow, non-linear or circular patterns of motility (VCL 39.05 ± 11.92; BCF 0.02 ± 0.35; LIN 5.15 ± 4.88). The recognition of 3 clearly identifiable subgroups supports our hypothesis that heterogeneity of sperm motility patterns exists within koala ejaculates. These may be a reflection of the heterogeneity in sperm-head morphotypes in koala semen, but that remains to be investigated in more detail. The clear distinctions between these groups, and the observation that all 3 subpopulations exist in each of the ejaculates, also suggest that the spermatozoa exhibit functional differences, possibly related to biochemical or maturational status. Many thanks to Dr. Michael Pyne and Dr. Vere Nicholson and their teams and animals at Currumbin Wildlife Sanctutary and Dreamwolrd QLD for all their help and support for the collection of samples.

scholarly journals Sperm motility activation in the critically endangered booroolong frog: the effect of medium osmolality and phosphodiesterase inhibitors

2017 ◽  
Vol 29 (11) ◽  
pp. 2277 ◽  
Author(s):  
Aimee J. Silla ◽  
Leesa M. Keogh ◽  
Phillip G. Byrne
Keyword(s):  
Sperm Motility ◽  
Phosphodiesterase Inhibitors ◽  
Critically Endangered ◽  
Computer Assisted ◽  
Sperm Activation ◽  
Sperm Analysis ◽  
Sperm Longevity ◽  
Pde Inhibitors ◽  
Activation Data ◽  
Optimal Procedures

Effective activation of sperm motility is fundamental to successful artificial fertilisation; however, studies investigating optimal procedures in amphibians are lacking. This study found the optimal osmolality of activation media for sperm motility activation and evaluated the effect of phosphodiesterase (PDE) inhibitors on sperm activation and longevity in the critically endangered booroolong frog, Litoria booroolongensis. To assess the effect of medium osmolality (10, 25, 50, 75, 100 and 200 mOsmol kg−1) and PDE inhibitors (control, 2.5 mM caffeine, 5 mM caffeine, 2.5 mM pentoxifylline, 5 mM pentoxifylline, 2.5 mM theophylline and 5 mM theophylline) on initial activation, percentage sperm motility and sperm velocity were quantified using computer-assisted sperm analysis. To assess the effect of PDE inhibitors (control, 2.5 mM caffeine and 2.5 mM theophylline) on sperm longevity, percentage motility and velocity were assessed hourly until 10 h after activation. High (>60%) percentage motility was achieved in a broad range of activation-medium osmolalities (10–75 mOsmol kg−1). PDE inhibitors did not have an effect on initial sperm motility or velocity, but caffeine and theophylline improved sperm longevity, significantly increasing motility and velocity at 8, 9 and 10 h after activation. Data also show that sperm longevity in L. booroolongensis is extreme, with spermatozoa remaining motile more than twice as long as those of any other anuran amphibian.

scholarly journals Assessment of Poverty by Municipalities in the Context of Population Ageing—The Case of Małopolskie Voivodeship

2021 ◽  
Vol 13 (5) ◽  
pp. 2563
Author(s):  
Małgorzata Ćwiek ◽  
Katarzyna Maj-Waśniowska ◽  
Katarzyna Stabryła-Chudzio
Keyword(s):  
The Elderly ◽  
Research Problem ◽  
Point Of View ◽  
Computer Assisted ◽  
Chi Square ◽  
Social Challenge ◽  
Depth Analysis ◽  
Qualitative Variables ◽  
Chi Square Test ◽  
Research Gap

This article undertakes the research problem of the assessment of the significance of poverty as a social challenge for local self-government units, and the differences in the assessment of the incidence of this phenomenon depending on the type of municipality. The authors also analyse the relationships between the ageing of the population and the assessment of the extent of poverty by municipalities. It must be pointed out that the undertaken problem has not been a subject of in-depth analysis thus far. Hence, this article fills the identified research gap in this field. The empirical part is based on the results of our own research, conducted using the Computer-Assisted Web Interview (CAWI) method on a sample of 144 municipalities of the Małopolskie Voivodship (Poland). In order to verify whether there is a relationship between the researched qualitative variables, the chi-square test of independence was used. In order to determine the relationships occurring between the categories of variables characterising the scale of the incidence of poverty and the remaining variables, a correspondence analysis was conducted. The research enabled us to find the issue of poverty to be one of the most important social problems from the point of view of municipalities. It is also worth noting that the degree of ageing in the population has an impact on the assessment of poverty among the elderly.

Comparison of Computer-assisted Sperm Motility Analysis Parameters in Semen from Belgian Blue and Holstein?Friesian Bulls

2007 ◽  
Vol 42 (2) ◽  
pp. 153-161 ◽  
Author(s):  
G Hoflack ◽  
G Opsomer ◽  
T Rijsselaere ◽  
A Van Soom ◽  
D Maes ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Holstein Friesian

Effect of separate and combined exposure of selenium and diazinon on rat sperm motility by computer assisted semen analysis

2016 ◽  
Vol 38 ◽  
pp. 144-149 ◽  
Author(s):  
Robert Toman ◽  
Svatoslav Hluchy ◽  
Michal Cabaj ◽  
Peter Massanyi ◽  
Shubhadeep Roychoudhury ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Combined Exposure

Impact of supplementing commercial rabbit pellets with fresh Moringa oleifera leaves on semen characteristics in rabbit bucks under tropical climate in Trinidad

2021 ◽  
Author(s):  
Krishna Mohan Kumar
Keyword(s):  
Sperm Motility ◽  
Moringa Oleifera ◽  
Sperm Count ◽  
Control Group ◽  
Computer Assisted ◽  
Group Iii ◽  
Group I ◽  
Significant Difference ◽  
Moringa Oleifera Leaves ◽  
The Impact

Objective This study aimed to evaluate the impact of the dietary supplement of Moringa oleifera leaves (MOL) on semen quality and characteristics in rabbits. Methods Eighteen (n=18) breeding bucks of New Zealand white, of similar age group, were used for the study. Three feeding regimes, (i) 100% commercial rabbit pellets (CRP)-Group I (ii) 90% CRP + 10% fresh MOL on a dry matter (DM) basis – Group II and (iii) 80% CRP + 20% fresh MOL on a DM basis – Group III, were adopted and the trial continued for 21 days. After adaptation to the diet, semen was collected from each buck and subjected to evaluation using a computer-assisted semen analyser. Results In Group III, the sperm count, normal sperm morphology, and sperm motility increased (52.0%) in comparison with the control (Group I; 50.1%). The inclusion of 20% Moringa oliefera in the diet (Group III) caused a significant increase (P<0.05) in semen concentration (Control =136.2 M/mL; Group III=297.2 M/mL). There was no significant difference (P>0.05) in sperm motility and semen volume among the groups. Conclusion The results suggest that supplementing commercial rabbit pellets with 20% fresh Moringa oliefera leaves on a DM basis can improve the quality and characteristics of semen in breeding bucks.

scholarly journals Simultaneous Determination of Local Cerebral Glucose Utilization and Blood Flow by Carbon-14 Double-Label Autoradiography: Method of Procedure and Validation Studies in the Rat

1986 ◽  
Vol 6 (3) ◽  
pp. 273-285 ◽  
Author(s):  
Myron D. Ginsberg ◽  
David W. Smith ◽  
Mitchell S. Wachtel ◽  
Mayra Gonzalez-Carvajal ◽  
Raul Busto
Keyword(s):  
Validation Studies ◽  
Optimization Procedure ◽  
Mean Value ◽  
Great Promise ◽  
Computer Assisted ◽  
Published Data ◽  
Carbon 14 ◽  
Coefficients Of Variation ◽  
Double Label ◽  
Label Strategy

Validation studies were undertaken to establish a computer-assisted double-label autoradiographic strategy employing [14C]2-deoxyglucose ([14C]2DG) and [14C]iodoantipyrine ([14C]IAP) to measure local CMRglu (LCMRglu) and CBF (LCBF). An organic solvent was used to extract the majority of IAP between first and second film exposures. In contrast to previously published data, all solvents tested produced partial losses of 2DG from tissue, and all allowed 2–6% of IAP to persist even after 5-day washes. Technical-grade chloroform permitted equal retention of unmetabolized and metabolized 2DG. A linear model was established, which was insensitive to the changes in tissue self-absorption that were shown to occur with chloroform extraction. Propagated error in computing tissue [14C]2DG and [14C]IAP was reduced by maximizing IAP extraction (by longer solvent wash times) and by administering 2.5 times as much IAP as 2DG. Fractional 2DG retention was measured in single-label 2DG sections placed on the films, and fractional IAP retention was evaluated by an optimization procedure. With this strategy, double-label values for LCMRglu and LCBF in anesthetized rats agreed with values obtained in matched single-label series to within 5%. The coefficients of variation for the double- and single-label LCMRglu data were virtually identical, whereas the coefficient of variation for double-label LCBF was 1.8 times that of single-label LCBF. The double-label strategy permitted pixel-by-pixel measurement and video display of the LCMRglu/LCBF ratio; the mean value among structures was 0.472 μmol/ml. With proper attention to methodological detail, this double-label strategy shows great promise for routine laboratory application.

scholarly journals Seminal and spermatic characteristics of fresh semen and the effects of sperm cooling in Steindachneridion melanodermatum (Garavello, 2005)

2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4493
Author(s):  
Ronan Maciel Marcos ◽  
Giovano Neumann ◽  
Cesar Pereira Rebechi de Toledo ◽  
João Marcos Sena ◽  
Gilmar Baumgartner ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Storage Temperature ◽  
Sperm Concentration ◽  
Plasma Osmolality ◽  
Storage Period ◽  
Sperm Velocity ◽  
Powdered Milk ◽  
Sperm Activation ◽  
And Storage ◽  
Fresh Semen

<p>This study describes the seminal and spermatic characteristics of fresh semen of <em>Steindachneridion melanodermatum </em>and investigates the effects of dilution, temperature, and storage period on its spermatic parameters. Sperm samples were collected from nine hormonally-induced males. The following parameters in fresh sperm were analyzed: seminal plasma osmolality (OSM), seminal pH, sperm motility (MOT), sperm velocity (SV) (including sperm curvilinear velocity (CVV), sperm straight-line velocity (SLV), and sperm average path velocity (APV)), total time of sperm motility (TEMP), sperm concentration (CONC), and index of sperm normality (NORM). Sperm samples from each male were diluted in a solution containing 5% fructose and 5% powdered milk, and stored at 10°C and 25°C. The same was carried out for sperm samples not subjected to dilution. From these samples, MOT, CVV, SLV, APV, SV, and TEMP were measured after 0 h, 5 h, 9 h, 18 h, 27 h, 36 h, 45 h, and 54 h. Males released 11.74 ± 5.38 mL of sperm, with an osmolality of 258.78 ± 29.36 mOsm.kg-1 and pH of 7.11 ± 0.31. The sperm presented a MOT of 99.86 ± 0.31% at a concentration of 1.03 × 1010 ± 3.65 × 109 spermatozoa.mL-1 with CVV of 185.58 ± 14.11 ?m.s-1, SLV of 49.15 ± 4.66 ?m.s-1, APV of 87.02 ± 4.13 ?m.s-1, SV of 106.52 ± 4.45 ?m.s-1, TEMP of 79.31 ± 5.62 s, NORM of 75.81 ± 5.71%. The results indicate that sperm motility, sperm velocity, and total time of sperm activation were affected by dilution, storage temperature, and storage period (p &lt; 0.05). Procedures for semen storage should be performed with undiluted sperm cooled at 10°C, or kept undiluted at 25°C for up to 27 h.</p>

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