Association of the cysteine-rich secretory protein-3 (CRISP-3) and some of its polymorphisms with the quality of cryopreserved stallion semen

2018 ◽  
Vol 30 (3) ◽  
pp. 563 ◽  
Author(s):  
Alexandra Usuga ◽  
Benjamín A. Rojano ◽  
Giovanni Restrepo

Contribution of seminal plasma proteins to semen freezability has been reported in several species, suggesting these proteins as genetic markers. The aim of this study was to evaluate the relationship between cysteine-rich secretory protein-3 (CRISP-3) and some of its single-nucleotide polymorphisms (SNPs) with post-thawing semen quality in stallions. DNA was obtained from 100 stallions, regions of interest were amplified by polymerase chain reaction and sequenced. Evaluated SNPs within the equine CRISP-3 gene were CRISP3c.+199A > G (SNP1), CRISP3c.+566C > A (SNP2), CRISP3c.+622G > A (SNP3) and CRISP3c.+716A > G (SNP4). CRISP-3 protein content in seminal plasma was determined by enzyme-linked immunosorbent assay. Semen from 30 stallions was cryopreserved and post-thaw motility, kinetics, abnormal morphology (AM), sperm vitality (SV) and membrane integrity (MI) were evaluated. Generalized linear models were fitted and means were compared using Tukey’s test. Correlation and regression analyses were performed. For SNP1 and SNP3, the AA genotype had the highest results for motility and MI; for SNP2, the best results for motility and AM were obtained with the CC genotype. For SNP4, the GG genotype had the lowest results, except for MI. A high level of CRISP-3 protein in seminal plasma had the best results for motility, kinetics, SV and AM. In conclusion, there was a relationship between CRISP-3 genotype and seminal plasma protein and post-thawing semen quality in stallions.

2013 ◽  
Vol 57 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Rafał Strzeżek ◽  
Krystyna Filipowicz ◽  
Marta Stańczak ◽  
Władysław Kordan

Abstract The resazurin reduction test (RRT) was subjected to spectrophotometric analysis to evaluate the quality of canine semen. Twenty four samples of canine semen were analysed. The absorption peaks for resazurin and resorufin were determined at 615 and 580 nm, respectively. The RRT ratio (RRTsperm-the ratio for samples containing spermatozoa, RRTplasma-the ratio for samples containing seminal plasma) was calculated by dividing the absorbance at 580 nm by the absorbance at 615 nm. Spearman’s correlation test was used to determine the significance of correlations between the analysed sperm parameters and the results of the resazurin reduction assay. The RRT ratio was highly correlated with sperm motility (r=0.68, P<0.01), progressive sperm motility (r=0.61, P<0.01), the subpopulation of cells with rapid velocity (r=0.72, P<0.01), and the subpopulation of cells with medium velocity (r= -0.54, P<0.05). A negative correlation was observed between the reducing capacity of seminal plasma vs. sperm with plasma membrane integrity (r= -0.60, P<0.01) and sperm with normal morphology (r= -0.58, P<0.01). The RRT test can be used as an additional tool for evaluation of the quality of canine semen.


2018 ◽  
Vol 33 (2) ◽  
pp. 62-70 ◽  
Author(s):  
A Hossain ◽  
MM Islam ◽  
F Naznin ◽  
RN Ferdousi ◽  
FY Bari ◽  
...  

Semen was collected from four rams, using artificial vagina and viability%, motility% and plasma membrane integrity% were measured. Fresh ejaculates (n = 32) were separated by modified swim-up separation using modified human tubal fluid medium. Four fractions of supernatant were collected at 15-minute intervals. The mean volume, mass activity, concentration, motility%, viability%, normal morphology and membrane integrity% (HOST +ve) of fresh semen were 1.0 ± 0.14, 4.1 ± 0.1 × 109 spermatozoa/ml, 85.0 ± 1.3, 89.4 ± 1.0, 85.5 ± 0.7, 84.7 ± 0.5 respectively. There was no significant (P>0.05) difference in fresh semen quality parameters between rams. The motility%, viability% and HOST +ve % of first, second, third and fourth fractions were 53.4 ± 0.5, 68.2 ± 0.3, 74.8 ± 0.3 and 65.5 ± 0.4; 55.5 ± 0.4, 66.2 ± 0.4, 74.5 ± 0.3 and 73.6 ± 0.3 and 66.7 ± 0.5, 66.8 ± 0.5, 65.2 ± 0.4 and 74.7 ± 0.5 respectively. The motility%, viability% and membrane integrity% of separated semen samples differed significantly (P<0.05) between four fractions. The mean motility% and viability% were significantly higher (P<0.05) in third fraction (74.8 ± 0.3%), whereas the mean HOST +ve% was significantly higher (P<0.05) in fourth fraction (74.7 ± 0.5). All quality parameters of separated spermatozoa were significantly (P<0.05) lower than that of fresh semen. The pregnancy rates were higher with fresh semen (71%) in comparison to that of separated sample (57%).Bangl. vet. 2016. Vol. 33, No. 2, 62-70


2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Maja Zakošek Pipan ◽  
Petra Zrimšek ◽  
Breda Jakovac Strajn ◽  
Katarina Pavšič Vrtač ◽  
Tanja Knific ◽  
...  

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.


Author(s):  
Ivan D. Sanchez-Diaz ◽  
Jesus Gonzalez-Feliu

This chapter studies the implication of aggregating establishments by categories with different levels of detail for modeling FTG. To this effect, the chapter conducts an assessment of freight trip generation (FTG) patterns homogeneity inside activity-based grouping. The method implemented is econometric in nature, which allows the assessment of the statistical significance of variables representing commercial activity sectors and sub-sectors. The results show that for some sectors the traditional high-level aggregation includes sub-sectors with homogenous FTG patterns and thus produces appropriate models; in some other cases (e.g., retail, manufacturing), the sub-sectors have different FTG patterns and thus more detailed data is needed to calibrate accurate models. This research can be used to enhance the efficiency of data collection, as it identifies some sub-sectors that need larger efforts for data collection, and some other categories where FTG homogeneity allows for less detailed data collection without hampering the quality of the models.


Author(s):  
Zhanhui Ou ◽  
Qirong Wen ◽  
Yu Deng ◽  
Yang Yu ◽  
Zhiheng Chen ◽  
...  

2015 ◽  
Vol 27 (1) ◽  
pp. 101

Stallions affected by seminal vesiculitis present history of infertility or subfertility, ejaculatory disturbance, spread of sexually transmitted pathogens, and changes in semen characteristics, leading to reduced semen quality and longevity. The aim of this study was to evaluate the semen quality of stallions with seminal vesiculitis before and after local treatment. Five stallions with a mean age of 12.4 years diagnosed with seminal vesiculitis were used. The identification of the microorganism involved in the pathogenesis of seminal vesiculitis of each animal was performed by bacterial culture of the seminal vesicles flush with Ringer Lactate solution, performed in duplicate at 1-week intervals. After identification of bacteria was performed, there was susceptibility testing to antibiotic (antibiogram) and the appropriate antibiotic was chosen. The local treatment was performed by endoscopy for 10 consecutive days, and this consisted of flushing with Ringer Lactate solution, followed by infusion of the antibiotic selected. The semen analyses were performed before starting the local treatment for seminal vesiculitis (M0), after a week (M1), and after a month (M2) of therapy. Sperm kinetics were performed by computerized method – CASA for the following parameters: percentage of sperm with total motility, progressive motility, and rapid sperm. Analysis of plasma membrane integrity was performed by epi-fluorescence microscopy, using the combination of fluorescent probes carboxyfluorescein diacetate and propidium iodide. Percentage of leukocytes was assessed through evaluation in light optical microscopy of semen smears stained with DiffQuick. The content of nitric oxide (NO) was determined by colourimetric Griess reaction by a spectrophotometer through the concentrations of nitrate (NO3–) and nitrite (NO2–). To perform the count of colony forming units per millilitre (CFU mL–1), an aliquot of 0.1 mL of semen was diluted in 9.9 mL of saline. A 0.1-mL aliquot of this sample was plated on Mueller-Hinton agar. The seeded plates were incubated, and the bacterial colonies were counted after 24 h. According to the performed dilution, total colonies identified corresponds to ×10 000 CFU mL–1. The data were analysed by two-way ANOVA followed by Tukey's test (P < 0.05). The values (mean ± standard error) of seminal parameters on M0, M1, and M2 were the following, respectively: sperm kinetics (total motility: 46.5 ± 5.13a; 75.1 ± 3.42b; 42.8 ± 5.28a; progressive motility: 19.3 ± 3.86a; 33.4 ± 2.39b; 16.5 ± 2.40a; rapid sperm: 22.2 ± 1.82a; 52.2 ± 5.65b; 22.1 ± 2.62a); plasma membrane integrity (47.5 ± 4.65a; 62.9 ± 5.41b; 39.1 ± 4.32a); percentage of leukocytes (35.2 ± 2.36a; 15.1 ± 2.55b; 36.1 ± 4.04a); CFU (119 980 × 103 ± 19 528.0 × 103a; 5375 × 103 ± 2453.7 × 103b; 65 850 × 103 ± 19 701.0 × 103ab) on fresh semen; and NO content (0.645 ± 0.172a, 0.117 ± 0.023b, 0.364 ± 0.110ab) on seminal plasma. The results demonstrate that local treatment after a week leads to an improvement in sperm quality; however, this was not maintained after 1 month of therapy, since the seminal parameters at this time are similar to pretreatment, which can be justified by recurrent disease.


2019 ◽  
Vol 22 (10) ◽  
pp. 882-889 ◽  
Author(s):  
María F García ◽  
Romina Nuñez Favre ◽  
María C Stornelli ◽  
Ramiro Rearte ◽  
María C García Mitacek ◽  
...  

Objectives The current study aimed to evaluate the relationship between specific seminal plasma components – cholesterol (CHOL), triacylglycerols (TAG) and total protein (PROT) concentrations – and semen quality in cats. A further aim was to determine the relationship between specific seminal protein bands and semen quality. Methods Thirteen toms, 2–5 years of age, were included. Semen collection was performed by electroejaculation every 4 weeks. Fifty-eight ejaculates were assessed for motility, velocity, volume, sperm concentration, total sperm count, viability, acrosome integrity, plasma membrane integrity and sperm morphology. Samples were divided into two groups: good semen quality (GSQ) and poor semen quality (PSQ). After evaluation, seminal plasma was separated from the sperm by centrifugation and stored at −20°C. CHOL, TAG and PROT concentrations were then assessed and seminal plasma protein profile was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results Seminal plasma CHOL and TAG concentrations, motility, velocity, sperm concentration, total sperm count and sperm morphology were significantly higher in GSQ cats compared with PSQ cats ( P <0.01). Moreover, seminal plasma SDS-PAGE analysis showed an identifiable extra band exclusively in the GSQ group. Conclusions and relevance Data obtained in this study showed that seminal plasma CHOL and TAG concentrations and specific protein bands could be used to improve semen evaluation in toms. In this sense, the 14 kDa protein band could be a valuable marker for semen quality in the cat and should be further investigated. However, more studies are necessary to determine its relationship with fertility.


1948 ◽  
Vol 38 (3) ◽  
pp. 323-331 ◽  
Author(s):  
T. Mann

SUMMARY1. A method is described whereby fructose content and fructolysis can be assayed accurately in small samples of semen. The advantages of this method lie in its simplicity, accuracy and practical convenience as a tool for the assessment of semen quality, applicable also under field conditions.2. The content of fructose in fresh semen depends upon the secretory function of accessory glands which is influenced directly by the activity of the male sex hormone. A low level of seminal fructose may coincide with other symptoms of hormonal malfunction and poor quality of spermatozoa. A high level of seminal fructose indicates satisfactory functional ability of the accessory glands, but it does not necessarily coincide with high quality of spermatozoa as expressed in terms of density and motility.3. The normal level of fructose in fresh semen undergoes frequent fluctuations which can be observed if semen collections are made from the same individual at different times. Considerable variations in the sperm/fructose ratio may also occur in the semen of the same individual as illustrated by the results of an ‘exhaustion test’.4. Fructose disappears from semen incubated in vitro. The rate of fructose disappearance forms a convenient measure of sperm fructolysis. The normal rate of fructolysis in bull semen is 1·4–2 mg. fructose per 109 sperm cells in 1 hr. at 37° C. At this high level it can be maintained until almost the whole reserve of fructose has been exhausted. Azoospermic and necrospennic semen, as well as that from vasectomized animals, are unable to utilize fructose. A reduced rate of fructolysis is found in low quality semen of subfertile and infertile animals.5. The conditions of sperm survival in semen incubated in narrow tubes as used for the fructolysis test as well as for storage of semen in the practice of artificial insemination, are almost purely anaerobic. Under such conditions the survival of spermatozoa must largely depend upon fructolysis and not upon respiration.


2020 ◽  
Author(s):  
Zhanhui Ou ◽  
Qirong Wen ◽  
Yu Deng ◽  
Yang Yu ◽  
Zhiheng Chen ◽  
...  

Abstract Purpose The effects of cigarette smoking on male semen quality are controversial, and the molecular mechanisms underlying how cigarette smoking affects semen quality are not clear yet. Methods In this study, semen samples from 70 heavy smokers and 75 non-smokers receiving infertility treatment were included. Basic semen parameters in non-smokers and heavy smokers were evaluated. Levels of glutathione (GSH), lipid reactive oxygen species (ROS), iron and GSH-dependent peroxidase 4 (GPX4) protein level) were observed in human seminal plasma and in GC-2Spd cells exposed to cigarette smoke condensate (CSC). Results Heavy smokers had significantly higher abnormalities (sperm viability and sperm progressive motility) than non-smoking counterparts. Comparing non-smokers group, GSH level was reduced in the group of heavy smokers (P<0.05). However, the level of lipid ROS and iron were significantly increased (P<0.05). Besides, GSH level was reduced following treatment with CSC for 24 h, while lipid ROS and iron levels were increased (P<0.05). However, the levels were reduced after being co-cultured with Ferrostatin-1 (Fer-1) (P<0.05). The level of GPX4 protein was reduced after being treated with CSC in 24 h, and increased after being co-cultured with Fer-1(P<0.05). Conclusion Cigarette smoking is associated with high level of ferroptosis in seminal plasma and affect semen quality.


Animals ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 414 ◽  
Author(s):  
Jiří Šichtař ◽  
Filipa Bubeníčková ◽  
Jitka Sirohi ◽  
Ondřej Šimoník

The aim of this study was to evaluate the effect of the addition of two types of seminal plasma (SP) after thawing on the functional characteristics of frozen–thawed (F–T) spermatozoa of poor freezing stallions during prolonged incubation periods. Seminal plasma from stallions with 35–40% (standard seminal plasma, (S-SP)) and 60–70% (above standard seminal plasma, (A-SP)) progressively motile spermatozoa after thawing was used. The motility, kinematic parameters (Computer Assisted Sperm Analysis), distribution of spermatozoa into subpopulations, integrity (carboxyfluorescein diacetate/propidium iodide staining), and functionality (hypo-osmotic swelling (HOS) test) of the spermatozoa plasma membrane were evaluated after thawing (T0) and after 30 min (T30) of incubation at 37 °C. There was no effect of SP addition on spermatozoa motility, but there was a significant positive effect on the kinematic parameters at T0 and T30. The addition of SP significantly increased the percentage of spermatozoa in the fast subpopulation at T0 as well as at T30. Plasma membrane integrity was not affected by the treatment, but functionality significantly decreased by 5% compared to the control group when samples were incubated for 30 min with A-SP. In conclusion, generally, the post-thaw addition of seminal plasma positively affected the post-thaw quality of semen from poor freezing stallions.


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