Estimation Of Sperm Motility Research Articles

Correlations Amongst Functional and Morphological Attributes and Oxidative Markers of Fresh and Cryopreserved Semen of Gir and Murrah Bulls

This study was undertaken during the winter season on healthy mature Gir cattle and Murrah buffalo bulls (n=3 each). The semen samples (6 ejaculates/bull, total 36 ejaculates) collected in the morning using artificial vagina were evaluated for routine seminal attributes, including acrosomal and plasma membrane integrity. The samples were then diluted @ 100 million sperm/ml with tris fructose yolk glycerol extender without and with sericin @ 0.1, 0.25, 0.5 and 1.0% (w/v), filled in French mini-straws, and frozen in LN2 using biofreezer as per standard freezing protocol. Straws were thawed in water bath at 37°C for 30 sec and evaluated for post-thaw quality, viz., motility, viability, morphology, acrosome integrity and plasma membrane integrity (HOST). Lipid peroxidation (malondialdehyde - MDA production) and activities of enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assessed as oxidative markers in seminal plasma of freshly diluted and frozen-thawed semen samples. Sericn at 0.5% level significantly (p less than 0.01) improved the post-thaw sperm quality with reduced oxidative stress in both the species. The breed-wise correlation coefficients (r) among sperm quality attributes and oxidative markers were studied in fresh and frozen-thawed semen of each species, and also for fresh with frozen-thawed semen. The findings revealed significant interrelationships amongst most of the attributes of fresh as well as post-thawed semen and also of fresh semen attributes with those of cryopreserved semen including oxidative markers in both the species. Sperm motility estimation in fresh, pre-freeze and post-thawed semen was a legitimately good indicator of quality of spermatozoa at various steps of semen processing/freezing, and its fertilizing potential. Thus, the sperm motility, HOS test and either MDA or SOD/GPx activity alone may be used as valuable and practical tools for routine assessment of bovine semen quality considering significant correlations found between them.

Computer assisted semen analysis for quantification of motion characteristics of bull sperm during cryopreservation cycle

Aim: The study was undertaken to quantify and to analyze the changes in the motion characteristics of bull spermatozoa during various stages of cryopreservation cycle. Materials and Methods: Using computer assisted semen analysis (CASA) technique, 26 ejaculates, collected from two Jersey bulls were analyzed for motility, head behaviour and swimming pattern of spermatozoa on dilution, pre-freeze and post-thaw stages of cryopreservation. French straw technique was employed for deep-freezing of semen using liquid nitrogen. O Results: Equilibration of diluted semen at 5 C has significantly (P< 0.01) reduced sperm motility, progressive motility, path velocity, and progressive velocity. Beat cross frequency was also affected significantly (P<0.05) by equilibration. Freezing and thawing processes drastically affected all the motility, velocity and head behaviour characteristics (P< 0.01). Conclusion: CASA facilitate objective evaluation sperm motion characteristics. Adoption of CASA technique has the potential for improvements in evaluation of semen thereby the quality of frozen semen for fertility can be enhanced. Artificial insemination (AI) in bovines is a classical assessment of quality of motility in terms of velocity, example for successful application of a biotech- swimming pattern, sperm head behaviour etc., may nological method at field level. Although fertility of help in better understanding of the possible sperm frozen-thawed semen is generally acceptable but the function. The advent of computer assisted semen analysis efficiency of cryopreservation is still relatively low. (CASA) has brought a new dimension to semen Rapid rate of cryopreservation results in cell damage evaluation. The CASA technique yields repeatable to spermatozoa due to osmotic imbalance (1). and highly reliable results on kinematics of ejaculates The mammalian spermatozoa acquire the capacity based on measurements of individual sperm cells. for flagellar movements during epididymal transport Therefore, assessment of sperm motility coupled with and it is modulated in the female reproductive tract kinetic measurements would help in better evaluation before termination during fertilization. These changes of semen quality. in flagellar activity reflect the physiological events within the sperm cell including the metabolic activity Materials and Methods and alteration of the sperm membrane (2). Further the Two Jersey bulls, aged 2 years, maintained for events of semen processing during cryopreservation instructional purpose for the students of Madras cycle bring injuries to sperm membrane leading to Veterinary College, Chennai under stall-fed conditions changes in the sperm physiology which may be were used for the study. The bulls were fed with 3 kg of reflected in sperm motility (3). Cryopreservation also concentrate/day/bull in addition to 3 kg of dry fodder affects the functional integrity of acrosome and per bull. Water and green fodder were provided ad mitochondria that is responsible for the generation of libitum. Since semen collection by artificial vagina energy from intracellular stores of ATP (4). method did not involve any intervention in the normal Conventionally, sperm motility estimation is done physiology of the animals, this experiment was by visual approximation of progressively moving exempted approval from Institutional Animal Ethics spermatozoa using phase contrast microscope. The

Evaluation of sperm fertilizing ability using the Sperm Quality Analyzer.

The Sperm Quality Analyzer is an inexpensive device which provides a quantitative estimation of sperm motility. To evaluate the fertilizing ability of human spermatozoa using a Sperm Quality Analyzer, correlations amongst the sperm motility index, the sperm penetration index (as assessed using the sperm penetration assay; SPA), and the fertilization rate in the treatment of standard IVF-ET were analysed retrospectively. The sperm motility index demonstrated a significant correlation with sperm concentration (p < 0.001), sperm motility (p < 0.001) and the motile sperm concentration (p < 0.001) in a total of 104 fresh semen samples from 81 men donating samples for IVF-ET. The sperm motility index also showed a significant correlation (p < 0.001) with the sperm penetration index in 60 patients, assessed using the SPA, before they were treated by standard IVF-ET. The correlation between the sperm motility index and the IVF-ET fertilization rate was higher than that between the sperm penetration index and the fertilization rate. The sperm motility index was classified into three categories: 'poor' (sperm motility index < 80), 'medium' (sperm motility index 81-160) and 'good' (sperm motility index > 160). The relationships between the IVF-ET fertilization rate and each category of the sperm motility index values were also evaluated. For the three categories in the sperm motility index, the fertilization rates (76.0%) of 60 samples judged as 'good' were significantly higher than those (44.2%) of 15 samples judged as 'medium' (p < 0.001) and those (34.7%) of 13 samples judged as 'poor' (p < 0.001). These results indicate that the Sperm Quality Analyzer provides a reliable estimation of the fertilizing ability of human spermatozoa.

Andrology: Density adjustment of software settings minimizes bias in automated sperm motility estimation

To minimize overestimation of motility, it is recommended that fresh semen be diluted with seminal plasma prior to automated analysis. However, for glycerolated or cryopreserved semen this is impractical, and alternative methods are needed to minimize automated motility bias. In the present study, the proportion of motile spermatozoa was determined in fresh, diluted and cryopreserved semen (n = 25 ejaculates) using visual and automated methods. The effect of software settings on motility was investigated by assessing samples at a range of modified settings. At standard settings, automated motility was biased in fresh semen (+7.2%) after dilution with cryopreservative (-2.9%) and after cryopreservation (-7.8%) (P < 0.0001 versus visual). Automated motility was inversely related to the minimum number of frames for motility sampling (P < 0.0001), with mean estimates of 41.0, 46.1, 52.0 and 58.2% generated at settings of 8, 4, 2 and 1 frame(s) respectively (n = 15 fresh, diluted and cryopreserved samples). Based on an arbitrary ordinal scale, a method was developed whereby motility sampling was adjusted prior to analysis according to sperm density. Analysis of an independent set of semen samples with density-adjusted software settings reduced bias in automated estimates (n = 30) before and after freezing (P < 0.0001). In addition, bias was no longer related to sperm density. In conclusion, modification of software settings is an effective alternative to dilution to minimize bias in automated motility estimates in fresh, diluted and cryopreserved human semen.

An assessment of sperm motility estimation for evaluation in rams

There is disagreement about the value of estimating spermatozoal motility as a predictor of fertility in rams. We tested the hypothesis that sperm motility can help one to predict lambing rate by evaluating sperm motility in rams prior to breeding and by subsequently breeding the rams regardless of their sperm motility results. A total of 943 rams (Merino, n=630 and Romney Marsh, n=313) was used during two breeding seasons. For fertility evaluation only 804 rams that bred at least ten ewes each were included in the study. Rams were grouped into 4 groups according to sperm motility scores, ranging from the lowest, which included rams with azoospermia, oligospermia, non-motile and single-motile sperm, through rams with 10 to 30%, 30 to 60%, to the highest at 60 to 100% motile sperm. The corresponding fertility rates found in our study in Merino and Romney breeds were 58.3 and 47.4%, 75.1 and 59.1%, 79.4 and 64.1% and 81.5 and 64.0%, respectively. We concluded that there was a statistically confirmed difference in fertility between the lowest and all the other motility categories. The differences in fertility among rams that had motility categories above 10% were not statistically significant. Evaluation of breeding soundness in large scale surveys of rams, based on sperm motility estimation, may be misleading. Clinical examination of genital organs seems to offer a reliable source of information about rams, and may be considered adequate in sheep breeding.

Semen characteristics in free-living koalas (Phascolarctos cinereus)

Spermic electroejaculates (range in motile sperm/ejaculate, 0.50-122.9 x 10(6); mean +/- s.e.m., 38.6 +/- 4.9) were recovered from 47 of 48 adult koalas captured from 3 wild populations in Australia. Semen was characterized by (i) a high density of globular bodies, which prevented the estimation of sperm motility without dilution; (ii) a brownish colour; and (iii) an acidic pH. Spermatozoa were categorized on the basis of 10 head forms, most cells being a curved or hooked shape. The koala populations differed in sperm concentration and motility ratings, but not in testes size, testosterone production or proportions of spermatozoa with various head shapes. These data confirm that free-living koalas normally produce spermatozoa with a high incidence of structural heterogeneity almost solely confined to the head region; and demonstrate the utility and safety of conventional gamete and endocrine studies, approaches which will be useful for determining the impact of genetic isolation and venereal disease on species fertility.

Determination of adenosine triphosphate in human semen to estimate the fertilizing potential and to quantify sperm antibodies

The measurement of the ATP content of fresh semen is as accurate as the estimation of sperm motility by conventional methods in discriminating between semen of fertile versus subfertile men. The ATP content of frozen thawed donor semen is correlated with the probability of conception per cycle of insemination. Exact quantification of cytotoxic sperm antibodies in serum is possible with the adenosine-triphosphate-release-cytotoxicity test, since measurement is free of the bias of microscopic examination. The procedure has been simplified by testing only one serum dilution and calculating the 'sperm toxicity index'.

Prospects and problems in developing a contraceptive vaccine for the male

There is disagreement about the value of estimating spermatozoal motility as a predictor of fertility in rams. We tested the hypothesis that sperm motility can help one to predict lambing rate by evaluating sperm motility in rams prior to breeding and by subsequently breeding the rams regardless of their sperm motility results. A total of 943 rams (Merino, n=630 and Romney Marsh, n=313) was used during two breeding seasons. For fertility evaluation only 804 rams that bred at least ten ewes each were included in the study. Rams were grouped into 4 groups according to sperm motility scores, ranging from the lowest, which included rams with azoospermia, oligospermia, non-motile and single-motile sperm, through rams with 10 to 30%, 30 to 60%, to the highest at 60 to 100% motile sperm. The corresponding fertility rates found in our study in Merino and Romney breeds were 58.3 and 47.4%, 75.1 and 59.1%, 79.4 and 64.1% and 81.5 and 64.0%, respectively. We concluded that there was a statistically confirmed difference in fertility between the lowest and all the other motility categories. The differences in fertility among rams that had motility categories above 10% were not statistically significant. Evaluation of breeding soundness in large scale surveys of rams, based on sperm motility estimation, may be misleading. Clinical examination of genital organs seems to offer a reliable source of information about rams, and may be considered adequate in sheep breeding.

Spermatozoal antigens — Alternative candidates for birth control vaccines

There is disagreement about the value of estimating spermatozoal motility as a predictor of fertility in rams. We tested the hypothesis that sperm motility can help one to predict lambing rate by evaluating sperm motility in rams prior to breeding and by subsequently breeding the rams regardless of their sperm motility results. A total of 943 rams (Merino, n=630 and Romney Marsh, n=313) was used during two breeding seasons. For fertility evaluation only 804 rams that bred at least ten ewes each were included in the study. Rams were grouped into 4 groups according to sperm motility scores, ranging from the lowest, which included rams with azoospermia, oligospermia, non-motile and single-motile sperm, through rams with 10 to 30%, 30 to 60%, to the highest at 60 to 100% motile sperm. The corresponding fertility rates found in our study in Merino and Romney breeds were 58.3 and 47.4%, 75.1 and 59.1%, 79.4 and 64.1% and 81.5 and 64.0%, respectively. We concluded that there was a statistically confirmed difference in fertility between the lowest and all the other motility categories. The differences in fertility among rams that had motility categories above 10% were not statistically significant. Evaluation of breeding soundness in large scale surveys of rams, based on sperm motility estimation, may be misleading. Clinical examination of genital organs seems to offer a reliable source of information about rams, and may be considered adequate in sheep breeding.

The estimation of sperm motility in semen, on a membrane slide, by measuring the area change frequency with an image analysing computer

A simple slide is described in which the base is a permeable membrane so that a suspension of spermatozoa (or other cells) may be examined under controlled conditions with a microscope. An objective method of assessing the motility of spermatozoa in undiluted or diluted semen from a wide variety of species including cattle, horse, pig, rabbit, rat and sheep is described. It is shown to be well correlated with other methods of assessing motility and also with the non-return rate obtained with frozen cattle semen.

The Thickness of Microscopically Examined Seminal Sample and its Relationship to Sperm Motility Estimation

Sperm motility was examined microscopically in 7 drops of different samples. By a special way of slide preparation it was possible to evaluate motility at various thickness of each drop, ranging from zero to 20 microns and at intervals of 1 micron. No sperm motility was found below 2 microns. Between 2 and 10 microns per cent and quality of motility as well as sperm maximal speed increased gradually. Above 10 microns all these values remained constant but vision was impaired as blurring of sperm appeared. Optimal conditions for obtaining reliable results of motility estimation therefore exists only at a narrow range of 8–10 microns of thickness. But without control, it is difficult to get this range. In most cases, motility is estimated where vision is clear but where friction may affect motility and the results are therefore of limited value. The implication of these facts on accuracy and conclusion of many studies on sperm motility done in the past are discussed.