199CHARACTERISTICS OF CHAMOIS (RUPICAPRA PYRENAICA PARVA) 
EPIDIDYMAL SPERMATOZOA DEPENDING ON TIME POSTMORTEM

The Spanish Cantabrian chamois (Rupicapra pyrenaica parva) is a wild ruminant of the Cantabric Mountains (North of Spain). It is not an endangered species, but it is vulnerable to sarcoptic mange outbreaks and it is appreciated as a hunting trophy. The aim of the present study, as part of a project for establishing genetic resource banks for wild species in the North of Spain, was to determine the effect of postmortem time (PT) on epididymal sperm quality. We obtained 37 sets of testes from males hunted during the breeding season (autumn). The samples, with testes inside of the scrotum to prevent drying, were cooled to 5°C, and processed at 4 different PTs (0–30h, 30–60h, 60–90h, 90–120h). Sperm was obtained from incisions made in the caudal epididymis. Osmolality (OSM) and pH of the undiluted fluid were measured with a cryoscopic osmometer (Osmomat-030, Gonotec TM; Berlin) and an electronic pH-meter (CG 837, SchottTM; Mainz, Germany), respectively. Sperm motility (M) and progressive motility (PM) were assessed subjectively at 37°C. An aliquot of sperm was fixed in glutaraldehyde and used to evaluate acrosome integrity (ACR) and abnormal forms (heads, AH; midpieces, AM; tails, AT). Membrane functionality (MF) was assessed by means of the HOS test (100mOsmkg−1, 18min). All analyses were carried out using a phase-contrast microscope (×100 for motility and ×400 for other analyses). We obtained the Spearman correlation coefficients between the analyzed parameters and PT. Samples were graded on sperm quality parameters (PM, ACR and MF: >60%, high; 60–30%, medium;<30%, low), and their distribution among PT groups was compared (χ2, P<0.05). The following parameters correlated significantly with PT (r and P are shown): pH (0.37, 0.008), OSM (0.52, <0.001), M (−0.51, <0.001), PM (−0.62, <0.001), MF (−0.44, 0.001), ACR (−0.33, 0.02), AT (0.41, 0.005). The correlations of pH and osmolality indicate changes in epididymal fluid composition which could impair sperm viability. In fact, sperm motility, acrosome integrity and membrane functionality showed negative correlations. Also, AT increased with PT, which could be related to membrane damage (along with MF decrease). The distribution of the samples in quality groups is shown in the Table 1. There were no low quality samples in the 0–30h group, but the proportion of such samples increased significantly with PT. In conclusion, epididymal sperm characteristics changed with PT, showing a remarkable loss of quality after the first 30h. Even so, it was still possible to find samples of acceptable quality after several days. This is an important observation when collecting samples for genome resource banking. However, it will be necessary to assess the fertilizing ability of sperm stored for extended periods postmortem to confirm the utility of these findings. Table 1 Variation of sperm quality (PM,ACR and MF: >60, high; 60%–30%, medium; <30%, low) with PT

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116 EFFECT OF INCUBATION OF BOVINE EPIDIDYMAL SPERMATOZOA IN SEMINAL PLASMA PRIOR TO CRYOPRESERVATION

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab116 ◽

2007 ◽

Vol 19 (1) ◽

pp. 175

Author(s):

C. A. Guerrero ◽

J. A. Jenkins ◽

J. W. Lynn ◽

K. R. Bondioli ◽

R. A. Godke

Keyword(s):

Seminal Plasma ◽

Sperm Quality ◽

Egg Yolk ◽

Quality Parameters ◽

Water Bath ◽

Sperm Chromatin ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Epididymal Sperm ◽

Acrosome Integrity

Studies examining the influence of seminal plasma on sperm function have shown both beneficial and detrimental effects. However, its effect on pre-frozen bovine epididymal sperm (BES) has not been documented. The objective of this study was to determine the effect of a 30-min incubation of BES in bovine seminal plasma (SEMP) prior to freezing on post-thaw sperm parameters. Paired testes were obtained from mature bulls (n = 10) at a local abattoir and transported to the laboratory (25–28°C) within 3–5 h postmortem. BES were harvested by multiple incisions from the caudae epididymides of each bull, pooled, and split into either Treatment A, 3 mL of egg yolk Tris-based medium (EYT) (No SEMP), or Treatment B, 3 mL of SEMP, each for 30 min in a 37°C water bath. SEMP was extracted from ejaculates of mature bulls (n = 6), and then pooled and stored at −20°C until used. Sperm from both treatments were re-suspended in EYT, adjusted to 70 × 106 sperm mL−1 and cooled to 4°C at 0.1°C min−1. Samples were diluted slowly over a 30-min period with 1:1 EYT medium containing 14% glycerol and loaded into 0.5-mL straws. Straws were frozen in liquid nitrogen vapors. For sperm analyses, straws were thawed in a water bath at 37°C for 40 s. Morphology was determined by staining with eosin-nigrosin. Viability and acrosome integrity were measured simultaneously with the combination of SYBR 14, propidium iodide, and PE-PNA. Mitochondrial activity was measured by the combination of MitoTracker Red and SYBR 14. DNA integrity was determined by acridine orange using the sperm chromatin structure assay (SCSA®; SCSA Diagnostics, Inc., Brookings, SD, USA). All assays were performed by multicolor flow cytometry. Differences between treatments were analyzed using one-way ANOVA (P < 0.05). In summary, all sperm quality parameters decreased significantly after thawing in both treatments (Table 1). A significantly higher overall and progressive motility post-thaw was achieved when sperm were incubated in SEMP prior to freezing. However, no difference was detected in sperm viability, acrosome integrity, mitochondrial activity, and DNA integrity between treatments. Also, SEMP reduced the quantity of distal droplets and broken tails post-thaw over that of no SEMP. Results indicate that incubation of BES in bovine seminal plasma prior to freezing improves post-thaw sperm quality. Table 1. Effect of bovine seminal plasma (SEMP) on quality parameters (mean ± SEM) of post-thaw bovine epididymal sperm

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Short communication: The percentage of egg yolk in the freezing media affects mouflon (Ovis musimon) epididymal sperm cryosurvival

Spanish Journal of Agricultural Research ◽

10.5424/sjar/2018163-13268 ◽

2018 ◽

Vol 16 (3) ◽

pp. e04SC04 ◽

Cited By ~ 1

Author(s):

Lucía Martínez-Fresneda ◽

Milagros C. Esteso ◽

Adolfo Toledano-Díaz ◽

Cristina Castaño ◽

Rosario Velázquez ◽

...

Keyword(s):

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Quality Parameters ◽

Epididymal Sperm ◽

Morphological Abnormalities ◽

Head Displacement ◽

Straight Line ◽

Acrosome Integrity ◽

Ovis Musimon

Sperm cryopreservation protocols are not well defined in many wild species such as the mouflon. The aim was to study the effect of different concentrations of EY on mouflon epididymal sperm cryosurvival. Samples were collected by the flushing method and cryopreserved by the conventional freezing technique in straws using a TEST (TES, Tris, glucose) 5% v/v glycerol medium containing either 6% v/v (n=16) or 12% v/v (n=13) clarified EY. The membrane integrity, acrosome integrity, motility, and morphological abnormalities were assessed in fresh and frozen/thawed samples. Fresh sperm quality parameters did not differ between groups except for the acrosome integrity that was lower in the TEST-6% EY than in the TEST-12% EY group (88.9 ± 2.1% vs 94.7 ± 0.8%). Membrane integrity (31.6 ± 4.6% vs 11.6 ± 4.5%), total motility (32.8 ± 4.6% vs 17.2 ± 5.6%), progressive motility (13.3 ± 2.7% vs 6.1 ± 2.9) were higher in frozen-thawed sperm with TEST-6% EY than with TEST-12% EY (p<0.05). Other motility parameters such as curvilinear velocity, straight-line velocity, average path velocity and amplitude of lateral head displacement were also higher (p<0.05) in frozen-thawed sperm with TEST-6% EY. Frozen-thawed acrosome integrity (85.1 ± 3.3% vs 91.9 ± 2.3) and morphological abnormalities (34.0 ± 3.7% vs 29.1 ± 3.6%) did not differ between extenders. In conclusion, high EY concentration had detrimental effects on post-thaw quality parameters, therefore the use of TEST based extender containing 6% EY is recommended for the cryopreservation of mouflon epididymal sperm.

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Antioxidant Effect of a Polyphenol-Rich Murtilla (Ugni molinae Turcz.) Extract and Its Effect on the Regulation of Metabolism in Refrigerated Boar Sperm

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/2917513 ◽

2019 ◽

Vol 2019 ◽

pp. 1-15 ◽

Cited By ~ 3

Author(s):

Ignacio Jofré ◽

Magdalena Cuevas ◽

Leticia Signori de Castro ◽

João Diego de Agostini Losano ◽

Mariana Andrade Torres ◽

...

Keyword(s):

Lipid Peroxidation ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Damage ◽

Antioxidant Effect ◽

Boar Sperm ◽

Fertilization Capacity ◽

Ugni Molinae

The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. We evaluated the effect of polyphenol-rich aqueous extract of murtilla (Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. Five experiments were performed: first, characterization of the polyphenol content from five genotypes of murtilla; second, determination of the genotype with the best antioxidant effect (MT-Ex); third, the antioxidant capacity on O2- and lipid peroxidation; fourth, the influence of MT-Ex on motility, calcium movement, cAMP, and metabolic parameters; and fifth, analysis of long-term refrigeration. The average phenolic content was 344 ppm; gallic acid, catechin, quercetin, myricetin, and kaempferol were detected. All extracts evaluated presented a concentration-dependent antioxidant effect. MT-Ex reduces intracellular O2-/peroxides but low lipid peroxidation. MT-Ex in nonstimulated ROS conditions reduces sperm motility, mitochondrial membrane potential, cAMP, and ATP, but the succinate dehydrogenase activity remained normal; also, we observed a reduction in calcium movement in in vitro sperm capacitation. The long-term analyses showed that MT-Ex improved sperm motility decay and reduced membrane damage and ROS at 168 h. Based on this study, we propose MT-Ex as a supplement in semen extenders.

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Sperm viability and morphology of two genetically diverse Merino lines

Reproduction Fertility and Development ◽

10.1071/rd99041 ◽

2000 ◽

Vol 12 (6) ◽

pp. 337 ◽

Cited By ~ 2

Author(s):

H. Lambrechts ◽

F. E. van Niekerk ◽

S. W. P. Cloete ◽

W. A. Coetzer ◽

G. van der Horst

Keyword(s):

Adverse Effect ◽

Sperm Motility ◽

Sperm Quality ◽

Sperm Parameters ◽

Sperm Viability ◽

Base Population ◽

Epididymal Sperm ◽

High Line ◽

Computer Aided

Microscopically evaluated sperm parameters, as well as computer-aided sperm motility analysis (CASMA), were used to assess sperm quality and the effect of cryopreservation on ram semen obtained from two genetically diverse Merino lines. These lines were divergently selected on maternal ranking values for multiple rearing ability from the same base population since 1986. Replacements in the high (+) line were preferentially the progeny of ewes rearing >1 lamb per joining. Progeny of ewes rearing <1 lamb per joining was preferred as replacements in the low (–) line. Sperm quality, as assessed by percentages of live, abnormal and acrosome-intact spermatozoa as well as by motility, was independent (P≤0.20) of line, time of sampling and their interaction in ejaculated samples obtained from the eight rams used as sires in 1995. Sperm quality of frozen–thawed samples was adversely affected (P≤0.01) by cryopreservation and thawing at 35˚C for 30 s relative to fresh ejaculated samples. No consistent differences between lines were found in epididymal sperm samples obtained from 12 slaughtered rams (6 from each line). The adverse effect (P≤0.05) of cryopreservation and thawing at 35˚C for 30 s on sperm viability and motility was also demonstrated for these samples.

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Implementing an open-access CASA software for the assessment of stallion sperm motility: Relationship with other sperm quality parameters

Animal Reproduction Science ◽

10.1016/j.anireprosci.2016.11.003 ◽

2017 ◽

Vol 176 ◽

pp. 11-19 ◽

Cited By ~ 24

Author(s):

Elisa Giaretta ◽

Mauro Munerato ◽

Marc Yeste ◽

Giovanna Galeati ◽

Marcella Spinaci ◽

...

Keyword(s):

Open Access ◽

Sperm Motility ◽

Sperm Quality ◽

Quality Parameters

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

THE INDIAN JOURNAL OF VETERINARY SCIENCES AND BIOTECHNOLOGY ◽

10.21887/ijvsbt.15.3.7 ◽

2020 ◽

Vol 15 (03) ◽

pp. 24-29

Author(s):

A. J. Dhami ◽

Tapasvi M Patel ◽

DV Chaudhari

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Winter Season ◽

Correlation Coefficients ◽

Plasma Membrane Integrity ◽

Murrah Buffalo ◽

Oxidative Markers

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.

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Effects of Taurine on Sperm Quality during Room Temperature Storage in Hu Sheep

Animals ◽

10.3390/ani11092725 ◽

2021 ◽

Vol 11 (9) ◽

pp. 2725

Author(s):

Liuming Zhang ◽

Yanhu Wang ◽

Tariq Sohail ◽

Yan Kang ◽

Haoyuan Niu ◽

...

Keyword(s):

Room Temperature ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Parameters ◽

Control Group ◽

Negative Effects ◽

Positive Effects ◽

Antioxidant Parameters ◽

Acrosome Integrity ◽

Hu Sheep

The present study aimed to investigate whether the presence of Tau protected Hu sheep sperm from ROS stress during storage at room temperature. The semen was diluted with extender (Tris-based) at room temperature, supplemented with different concentrations of Tau (0, 10, 20, 40, 80, or 100 mM), and stored at 15 °C. Sperm quality parameters (sperm progressive motility, kinetic parameters, plasma membrane integrity rate, acrosome integrity rate, and MMP) and antioxidant parameters (ROS, MDA, SOD, CAT, and T-AOC) were evaluated during the preservation of semen. The addition of Tau, especially at a concentration of 20 mM, exerted positive effects on sperm quality parameters and antioxidant parameters compared to the sperm without Tau treatment (control group). The addition of Tau, especially at a concentration of 100 mM, exerted negative effects on sperm quality parameters and antioxidant parameters compared to the control group. Interestingly, the results indicated that the sperm acrosome integrity rate did not change during storage time. In conclusion, the addition of Tau to sperm preserved at room temperature can enhance the antioxidant ability of sperm, reduce the LPO on the 5th day, and improve the quality of semen preserved at room temperature. These results implied that Tau had potential to enhance Hu sheep sperm reproductive performance.

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CASA-Mot in mammals: an update

Reproduction Fertility and Development ◽

10.1071/rd17432 ◽

2018 ◽

Vol 30 (6) ◽

pp. 799 ◽

Cited By ~ 16

Author(s):

J. L. Yániz ◽

M. A. Silvestre ◽

P. Santolaria ◽

C. Soler

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Epidemiological Survey ◽

Quality Parameters ◽

Farm Animals ◽

Control Analysis ◽

Precise Calculation ◽

Sources Of Variation ◽

Basic Studies ◽

Physical And Chemical

Sperm motility is one of the most widely used parameters of sperm quality. Computer-aided sperm motility analysis (CASA-Mot) systems were developed to reduce the subjectivity of sperm motility assessment, and have had broad scientific and practical acceptance. In this review, the sources of variation and current applications of this technology and its relationships with other sperm quality tests are described in detail. Despite remarkable advances in the technique, there is still great need for standardisation in many species, and the numerous factors that affect the results make it difficult to provide universally accepted criteria for classifying semen samples based on sperm motility characteristics. The main fields for CASA-Mot include the study of male fertility and pathologies, evaluation of the effects of physical and chemical agents, improvement of epidemiological survey studies, more precise calculation of seminal doses for farm animals, realisation of basic studies about sperm function, improvement of sperm technologies such as cryopreservation and quality control analysis. Numerous relationships have been established between CASA-Mot and other sperm quality tests, although most of these parameters are complementary. Future CASA-Mot systems will probably be able to integrate several sperm quality parameters with motility.

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Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2016-0019 ◽

2016 ◽

Vol 19 (1) ◽

pp. 147-158 ◽

Cited By ~ 2

Author(s):

M. Lecewicz ◽

W. Kordan ◽

A. Majewska ◽

S. Kamiński ◽

A. Dziekońska ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Platelet Activating Factor ◽

Quality Parameters ◽

Sperm Viability ◽

Plasma Membrane Integrity ◽

Positive Effects ◽

Bull Semen

Abstract The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M. The experiment demonstrated that PAF at concentration 1×10-9M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10-9M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10-9M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10-9M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.

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Exogenous Oleic Acid and Palmitic Acid Improve Boar Sperm Motility via Enhancing Mitochondrial Β-Oxidation for ATP Generation

Animals ◽

10.3390/ani10040591 ◽

2020 ◽

Vol 10 (4) ◽

pp. 591 ◽

Cited By ~ 2

Author(s):

Zhendong Zhu ◽

Rongnan Li ◽

Chengwen Feng ◽

Ruifang Liu ◽

Yi Zheng ◽

...

Keyword(s):

Oleic Acid ◽

Palmitic Acid ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Female Reproductive Tract ◽

Positive Effects ◽

Boar Sperm ◽

Atp Generation ◽

Acrosome Integrity

It takes several hours for mammalian sperm to migrate from the ejaculation or insemination site to the fertilization site in the female reproductive tract in which glucose, amino acids, and fatty acids are regarded as the primary substrates for ATP generation. The present study was designed to investigate whether oleic acid and palmitic acid were beneficial to boar sperm in vitro; and if yes, to elucidate the mechanism that regulates sperm motility. Therefore, the levels of oleic acid and palmitic acid, motility, membrane integrity, acrosome integrity, and apoptosis of sperm were evaluated. Moreover, the enzymes involved in mitochondrial β-oxidation (CPT1: carnitine palmitoyltransferase 1; ACADVL: long-chain acyl-coenzyme A dehydrogenase) were detected with immunofluorescence and Western blotting. Consequently, the ATP content and the activities of CPT1, ACADVL, malate dehydrogenase (MDH), and succinate dehydrogenase (SDH) were also measured. We observed that CPT1 and ACADVL were expressed in boar sperm and localized in the midpiece. The levels of oleic acid and palmitic acid were decreased during storage at 17 °C. The addition of oleic acid and palmitic acid significantly increased sperm motility, progressive motility, straight-line velocity (VSL), membrane integrity, and acrosome integrity with a simultaneous decrease in sperm apoptosis after seven days during storage. When sperm were incubated with oleic acid and palmitic acid at 37 °C for 3 h, the activities of CPT1 and ACADVL, the ATP level, the mitochondrial membrane potential, the activities of MDH and SDH, as well as sperm motility patterns were significantly increased compared to the control (p < 0.05). Moreover, the addition of etomoxir to the diluted medium in the presence of either oleic acid or palmitic acid and the positive effects of oleic acid and palmitic acid were counteracted. Together, these data suggest that boar sperm might utilize oleic acid and palmitic acid as energy substrates for ATP production via β-oxidation. The addition of these acids could improve sperm quality.

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