Collection and evaluation of semen collected from jaguarundi (Puma yagouaroundi) through urethral catheterization and electroejaculation (original) (raw)
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Journal of Andrology, 2009
Assessing semen quality is crucially important for the exploitation of genetically superior sires in an artificial insemination (AI) program. In this study, we compare modern and conventional techniques to estimate bovine sperm concentration and membrane integrity. First, the NucleoCounter SP-100 was validated for sperm concentration and provided statistically reliable and repeatable estimates among aliquots and replicates of 25 fresh ejaculates. Sperm concentrations in 78 ejaculates were then determined with hemacytometer, flow cytometer, and NucleoCounter SP-100 and were significantly correlated (P , .001), with regression coefficients among these 3 techniques close to 1 (P , .01). However, the sperm concentration determined by hemacytometer was lower (P , .01) than by flow cytometer and NucleoCounter SP-100. Forty frozenthawed semen samples were then assessed for sperm concentration and membrane integrity with hemacytometer, flow cytometer and NucleoCounter SP-100. Significant relationships were found for sperm concentration determined by hemacytometer and Nucleo-Counter SP-100 and for sperm membrane integrity determined by flow cytometer and NucleoCounter SP-100 (P , .01). Finally, the standard curves of sperm concentrations in 6 spectrophotometers, comparing optical density against counts drawn by hemacytometer and NucleoCounter SP-100 (n 5 94 fresh ejaculates) showed different (P , .01) intercepts and regression coefficients (linear, quadratic, cubic). It was calculated that a breeding station can improve its production potential by 13% with the use of Nucleo-Counter SP-100 instead of hemacytometer for calibration of spectrophotometers. Flow cytometer and NucleoCounter SP-100 can be used with equal confidence to estimate sperm concentration and membrane integrity in domestic animals and human semen.
Semen quality assessments and their significance in reproductive technology
2013
Semen quality assessment methods are very important in predicting the fertilizing ability of persevered spermatozoa and to improve animal reproductive technology. This review discusses some of the current laboratory methods used for semen quality assessments, with references to their relevance in the evaluation of male fertility and semen preservation technologies. Semen quality assessment methods include sperm motility evaluations, analyzed with the computer-assisted semen analysis (CASA) system, and plasma membrane integrity evaluations using fluorescent stains, such as Hoechst 33258 (H33258), SYBR-14, propidium iodide (PI), ethidium homodimer (EthD) and 6-carboxyfluorescein diacetate (CFDA), and biochemical tests, such as the measurement of malondialdehyde (MDA) level. This review addresses the significance of specific fluorochromes and ATP measurements for the evaluation of the sperm mitochondrial status. Laboratory methods used for the evaluation of chromatin status, DNA integrity, and apoptotic changes in spermatozoa have been discussed. Special emphasis has been focused on the application of proteomic techniques, such as two-dimensional (2-D) gel electrophoresis and liquid chromatography mass spectrometry (LC-MS/MS), for the identification of the properties and functions of seminal plasma proteins in order to define their role in the fertilization-related processes.
Effectiveness of the sperm quality analyzer (SQA-Vp) for porcine semen analysis
Theriogenology, 2011
The Sperm Quality Analyzer (SQA-Vp) was evaluated for assessing concentration and motility of porcine semen. Both fresh and diluted semen from 50 different boars from a commercial artificial insemination (AI) centre were investigated. For the fresh ejaculate, the concentration obtained with SQA-Vp was compared with a photometer and a haemocytometer. For the diluted samples, the concentration and motility were compared with computer assisted semen analysis (CASA) and visual sperm analysis. The agreement between methods was studied with Bland-Altman plots and the repeatability with coefficient of variation (CV) as well as Bland-Altman plots. The sperm concentration (x106/ml) obtained with SQA-Vp (379.3 ± 134.9) for fresh ejaculates agreed well with concentration by the photometer (447.2 ± 154.2; difference= -67.9 x 106/ml; difference + 2SD = 55.3 x 106/ml; difference – 2SD = -191.1 x 106/ml) and with the haemocytometer (332.8 ± 141.11; d = 92.8; d + 2SD = 448.6; d – 2SD = -263). For diluted semen, the agreement between the concentration (x106/ml) assessed with SQA-Vp (20.4 ± 4.3) was good with CASA (23.2 ± 5.8; d = -2.8; d + 2SD = 6.2; d – 2 SD = -11.8) but poor with the haemocytometer (18.8 ± 5.0; d = 1.6; d+ 2SD = 12.2; d – 2SD = -9). The % motile spermatozoa assessed by SQA-Vp (65.8 ± 10.0) in diluted semen agreed well with CASA (72.2 ± 13.7; d = -6.4; d+ 2SD = 20; d – 2SD = -32.8) and with visual assessment (64.1 ± 11.6; d = 1.7; d+ 2SD = 30.9; d – 2SD = -27.5). The SQA-Vp showed a good repeatability (CV; repeatability coefficient) for measuring the concentration of both fresh (3.9%; d = 10.7; d + 2SD = 30.9; d - 2SD = -9.5) and diluted semen (2.6%; d = 1.0; d + 2SD = 2.38; d - 2SD = -0.42) and for motility (3.2%; d = 0.9; d + 2SD = 8.5; d - 2SD = -6.7). The mean SQA-Vp values fell between the other methods′ results for both fresh and diluted semen. Moreover the repeatability was acceptable. Therefore SQA-Vp can be used as a valid device for sperm quality analysis in pigs.
This study is part of an ongoing project on artificial insemination in ostriches. The physical output of neat semen from four ostrich males was investigated and the effect of reconstituting semen with: 1) seminal plasma of the same male (SPS); 2) seminal plasma of another male (SPD), and 3) Dulbecco's Modified Eagles Medium (DMEM). Semen was collected daily from one or two pairs of males using the dummy female method, each pair being replicated twice. Spermatozoa viability in neat semen, SPS, SPD and DMEM was assessed using nigrosin-eosin staining and the proportions of live normal, live abnormal and dead sperm were determined. Semen volume (mean ± SE) was 1.27 ± 0.13 mL, the concentration of spermatozoa 3.68 ± 0.17 x 10 9 /mL and the number of spermatozoa 4.92 ± 0.64 x 10 9 /ejaculate. Furthermore, the live normal, live abnormal and dead spermatozoa in the neat semen were 61.2 ± 4.5%, 21.2 ± 2.7% and 17.7 ± 4.3% respectively. The ejaculate volume and the number of dead spermatozoa were not affected by collection time. However, the number of live abnormal spermatozoa increased through the day causing a reduction in live normal spermatozoa. Furthermore, re-suspending spermatozoa in DMEM reduced the number of live normal (31.4 ± 4.6%) and live abnormal spermatozoa (11.0 ± 2.7%) and increased the number of dead spermatozoa (57.6 ± 4.4%). In contrast, numbers of live spermatozoa were higher when suspended in seminal plasma and similar in SPS (53.9 ± 4.6%) and SPD (50.7 ± 4.6%). These are the first crucial steps to determining the optimum semen collection time and to improving the viability of diluted spermatozoa.
Validation and Usefulness of the Sperm Quality Analyzer (SQA II-C) for Bull Semen Analysis
Reproduction in Domestic Animals, 2005
In this study, an upgrade version of the Sperm Quality Analyzer (SQA), the SQA-IIC was tested for the assessment of bull semen quality. In Expt 1, the device showed good repeatability of measurements within and between capillaries, as evidenced by the low coefficients of variation (CVs; <13%) at concentrations between 35 and 705 • 10 6 spermatozoa/ml. In Expt 2, 10 semen concentrations (1-1000 • 10 6 /ml) were stored in HEPES TALP for 48 h at room temperature. A timedependent decrease in sperm motility index (SMI) values was noticed. SMI values increased linearly with increasing sperm concentrations, but remained constant around 500, corresponding to a concentration of approximately 50 • 10 6 /ml. For sperm concentrations below 50 • 10 6 /ml, SMI values were highly correlated with concentration (p < 0.05) and with semen parameters, expressing the overall semen quality (p < 0.05; Expt 3). In Expt 4, a correlation of only 0.44 (p < 0.05) between SMI values of frozen-thawed semen samples of 35 bulls and the corrected 56-day non-return rate (56dNRRc) was found. Prediction of the 56dNRRc based on the SMI value of a semen sample was inaccurate. The present study indicates that the SQA-IIC is suitable for a rapid screening of bull semen diluted to a concentration of approximately 50 • 10 6 /ml. Furthermore, the device seems inappropriate for fertility prediction.
Application of semen evaluation techniques
Acta Agraria Debreceniensis, 2014
Laboratory methods of semen evaluation are used to select males for artificial insemination. The current review describes several techniques that have been recently used for sperm analysis. Conventional microscopic methods in combination with the objective computerassisted sperm motility and morphology analyzers and flow cytometry, allows to obtain more precise information about the membrane and functional status of spermatozoa. By using several methods to detect motility, viability, acrosomal and capacitation status besides DNA integrity sperm biology and some of the mechanism involved in sperm cry injury can be better understood. The number of possible targets related to sperm quality is increasing, and possible that some of them could enable sperm analysis for predicting freezability and fertility to be improved.
Animals, 2020
The guinea pig, as a livestock species, is still developing and growing throughout Peru and neighboring countries, as reflected by its increasing export since 2000. However, the selection of proven fertile males is tedious due to the absence of seminal parameter standards and the lack of safe semen collection techniques. Thus, pregnancy detection or live births are required for males’ selection. The purpose of this study was to describe the qualitative and quantitative semen parameters of fertile guinea pig males, to set reference values, and to validate a novel electroejaculation technique for the species. Semen was collected at weekly intervals from sixteen fertile males. Four transrectal electroejaculations were performed per male with 95% successful collections, yielding 39 viable semen samples. Seminal characteristics were as follows: pH 7.0 ± 0.13; ejaculate volume 0.67 ± 0.55 mL; sperm motility 90.81 ± 6.64%; sperm concentration 36.7 ± 28.41 × 106 sperm/mL; sperm count 20.09 ...
Theriogenology, 2006
Epididymal semen is being more often considered as a potential source of valuable genes for genome resource banks. To utilize this resource as efficiently as possible, storage and freezing fertility and preservation characteristics of epididymal semen have to be examined. Because semen quality should be assessed as objectively as possible, we introduced computer assisted sperm analysis (CASA) of epididymal bull semen. The aims of this study were: to determine the quality of fresh cauda epididymal bull sperm, conventionally and by CASA (Hamilton-Thorne Ceros 12.1); to compare epididymal sperm movement with the motion characteristics of ejaculated semen; and to investigate whether equality of semen characteristics exists between both caudae epididymides of the same bull. In experiment 1, it is shown that epididymal sperm has a lower motility (total: 48.7% versus 79.9%, p < 0.0001 and progressive: 34.4% versus 58.4%, p < 0.0001) and moves less straight (80.5% versus 84.5%, p < 0.0009) with a higher amplitude (6.1 mm versus 5.0 mm, p < 0.0001) than ejaculated semen. The epididymal straight line velocity (85.2 mm/s versus 98.3 mm/s, p < 0.0001) is lower, but the curvilinear velocity (173.5 mm/s versus 156.4 mm/s, p < 0.0001) is higher than those of ejaculated semen. The data in experiment 2 are analysed to determine equality, rather than to find a difference. They illustrate that mean differences, for most semen parameters, between the semen from paired caudae epididymides, deviated more than 20% from the average values of these parameters from all bulls; the exceptions (those parameters within 20% of the average for all bulls) were the percentage of live spermatozoa, the linearity of sperm movement, the weights of testis and epididymis, the weights of the cauda epididymis alone, the volumes, and the amplitudes of movement of the semen ( p < 0.05). The mean differences between the percentage of live spermatozoa and the amplitude of movement of the epididymal semen of both epididymides of one bull, were the only values smaller than 10% of the average value of this parameter ( p < 0.05). This implies that sperm from one cauda epididymis should not be used as a control for the other because, for most of the semen parameters (concentration, morphology, motility, and beat cross frequency), equality between caudae epididymides of the same bull could not be established. #
Comparison of Two Different Method for Sperm Concentration Measurement of Ram and Buck Semen
2012
Comparison of Two Different Method for Sperm Concentration Measurement of Ram and Buck Semen R Iis Arifiantini, Ririn Riyanti and WM Nalley Department of Veterinary Clinic, Reproduction and Pathology, Faculty of Veterinary Medicine, Bogor Agricultural University, Indonesia, Internship Student, Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor, Indonesia, Faculty of Animal Science, University of Nusa Cendana Kupang, Indonesia Corresponding email: iis.arifiantinipurna@gmail.com