The Prediction of Sperm Chromatin Structure Assay (SCSA) Parameters for Outcomes of Intrauterine Insemination

2020 ◽  
Author(s):  
Jiangman Gao ◽  
Liying Yan ◽  
Ying Huang ◽  
Hui Jiang ◽  
Yuanyuan Wang ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Pregnancy Loss ◽  
Loss Rate ◽  
Intrauterine Insemination ◽  
Sperm Concentration ◽  
Clinical Pregnancy ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Female Age ◽  
Sperm Dna

Abstract Background: Many studies have assessed the association between sperm DNA fragmentation and outcomes of ART. But the published papers have not offered enough evidence about whether sperm DNA fragmentation tests could make suggestions to predict intrauterine insemination (IUI) outcomes. The aim of this study was to assess whether the sperm chromatin structure assay (SCSA) parameters, sperm DNA fragmentation index (DFI) and high DNA stainability (HDS), could be used as predictors for treatment outcomes in IUI program.Methods: A retrospective cohort study was conducted at a large reproductive medicine center. 1139 IUI cycles from 1139 couples were studied. The association of SCSA parameters with the clinical pregnancy and early pregnancy loss after IUI were analyzed. Results: Clinical pregnancy rate per cycle in DFI<15%, 15%≤DFI<30%, and DFI≥30% groups were 11.1%, 7.3%, and 8.9%, respectively, with no statistical differences between the groups (P=0.127). Pregnancy loss rate were 24.3%, 27.6%, and 14.3%, respectively, with no statistical differences (P=0.762). Clinical pregnancy rate per cycle in HDS≤15% and HDS>15% groups were 9.8% and 7.5%, respectively, with no significant difference (P=0.468), and the pregnancy loss rate were 26.2% and 0, respectively, and also no statistical difference (P=0.191). Multivariate logistic regression analysis showed a higher rate of clinical pregnancy in couples with a younger female (OR=0.90, 95% CI: 0.83-0.97, P=0.007), and in couples with a male who had higher sperm concentration after washing (OR=1.02, 95% CI: 1.00-1.04, P=0.035). A higher risk of pregnancy loss was observed with increased female age (OR= 1.43, 95% CI:1.09-1.89, P=0.010) and lower sperm concentration after washing (OR= 0.92, 95% CI: 0.84-0.99, P= 0.029). Conclusions: Sperm DFI and HDS were not significantly correlated with clinical pregnancy and pregnancy loss in cycles of IUI. Female age and motile sperm concentration had statistically significant effects on both clinical pregnancy and pregnancy loss after IUI treatment.

313 ASSESSMENT OF SPERM DNA FRAGMENTATION IN CANINE EJACULATES USING THE Sperm-Halomax® KIT: PRELIMINARY RESULTS

2010 ◽  
Vol 22 (1) ◽  
pp. 312 ◽  
Author(s):  
M. Hidalgo ◽  
M. R. Murabito ◽  
M. J. Gálvez ◽  
S. Demyda ◽  
L. J. De Luca ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Sperm Concentration ◽  
Semen Parameters ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Commercial Kit ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

Recently, a new procedure for the analysis of sperm DNA fragmentation has been developed for humans and different mammalian species, using a commercial kit based on the sperm chromatin dispersion (SCD) test; however, a descriptive study in canine semen has not been performed. The aim of this work was to assess the sperm DNA fragmentation in canine ejaculates using the SCD test and 2 different staining techniques. For this purpose, ejaculates were collectedby digital manipulation from4 healthy dogs of different breeds (1 German Pointer, 2 Spanish Greyhounds, and 1 Crossbreed). After collection, the sperm-rich fraction of the ejaculates from 3 dogs were pooled each time (n = 4) and then extended in Dulbecco’s phosphate buffered saline. All the pooled semen samples presented physiological values concerning routine semen parameters (motility, morphology, and sperm concentration). The sperm DNA fragmentation was assessed using the Sperm-Halomax® commercial kit specifically developed for canine semen (Halotech DNA SL, Madrid, Spain). Two semen aliquots of the diluted pooled semen samples were processed on each pre-treated slide provided in the kit following the manufacturer’s instructions. The last step was the staining technique. We stained each slide with 2 different staining procedures. The first half of the slide was stained with propidium iodide (Sigma-Aldrich, St. Louis, MO, USA) mixed in a proportion 1 : 1 with an antifading solution. The second half of the slide was stained for 15 min in Wright solution (1.01383.0500, Merck, Whitehouse Station, NJ, USA) 1 :1 in Phosphate Buffer pH 6.88 (1.07294.1000, Merck). The stained slides were observed using fluorescence and light microscopy, respectively. Five hundred sperm per slide were counted. Spermatozoa with fragmented DNA showed a large and spotty halo of chromatin dispersion. Unfragmented sperm only showed a small and compact halo. Statistical analyses were performed using the Statistical Package for Social Science version 12.0 (SPSS Inc., Chicago, IL, USA). The sperm DNA fragmentation index was compared between Wright and fluorescence staining methods by ANOVA. Results were expressed as mean ± standard error of the mean. The first report of the sperm DNA fragmentation index in canine ejaculates was 2.26 ± 0.53% for Wright staining and 1.99 ± 0.10% for fluorescence technique. No differences were found between staining procedures. In conclusion, it was possible to assess the sperm DNA fragmentation of canine ejaculates using 2 different staining procedures, expecting that continuous research could be useful in defining the role of DNA fragmentation SCD test in canine semen evaluation and cryopreservation.

scholarly journals Sperm DNA Fragmentation Among Smokers and Non-Smokers as Measured by Sperm Chromatin Dispersion Test

2018 ◽  
Vol 17 (1) ◽  
Author(s):  
Azha Syahril Azizan ◽  
Kamarul Bahyah Mustafa ◽  
Azantee Yazmie Abdul Wahab ◽  
Roszaman Ramli ◽  
Nurkhairulnisa Abu Ishak
Keyword(s):  
Dna Fragmentation ◽  
Seminal Fluid ◽  
Sperm Concentration ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Cross Sectional ◽  
Fluid Analysis ◽  
Sperm Dna ◽  
Dispersion Test ◽  
Sperm Chromatin Dispersion Test

Introduction: Traditionally, seminal fluid analysis is done to assess male infertility particularly sperm concentration, morphology and motility. Sperm chromatin dispersion test explores the quality of the sperm DNA structures. High sperm DNA fragmentation is associated with unexplained infertility, failure of assisted reproduction and recurrent miscarriages. Smoking has been associated with high sperm DNA fragmentation in some studies but not in others. Materials and Methods: A comparative cross-sectional study, involving 32 smokers and 32 non-smokers to determine the association between cigarette smoking and sperm DNA fragmentation. Semen samples were collected from patients undergoing seminal fluid analysis (SFA) at the IIUM Fertility Centre and IIUM Medical Centre O&G Clinic from January 2017 to June 2018. Sperm chromatin dispersion test was done and level of 15% is considered upper limit of normal. Results: 53.1% in the smoker group have abnormal level of sperm DNA fragmentation compared to 34.4% of nonsmokers. The median and interquartile range for smokers were 15.30 and 19.0 (10.425 - 29.375) respectively while for non-smokers were 8.25 and 17.8 (4.075 - 21.850) which was statistically significant (p= 0.012). It was also noted that three participants in the smoker group (9.4%) has very high sperm DNA fragmentation index of more than 80% even though they have normal seminal fluid analysis. Conclusion: There is an association between smoking and high sperm DNA fragmentation. There is a trend of increased in DNA fragmentation in smokers even though their SFA results were normal.

Sperm DNA fragmentation measured by sperm chromatin dispersion impacts morphokinetic parameters, fertilization rate and blastocyst quality in ICSI treatments

Zygote ◽  
2021 ◽  
pp. 1-8
Author(s):  
Shikai Wang ◽  
Weihong Tan ◽  
Yueyue Huang ◽  
Xianbao Mao ◽  
Zhengda Li ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Embryo Quality ◽  
Fertilization Rate ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Blastocyst Formation ◽  
High Quality ◽  
Morphokinetic Parameters ◽  
Sperm Dna ◽  
Blastocyst Quality

Summary To determine the effects of sperm DNA fragmentation (SDF) on embryo morphokinetic parameters, cleavage patterns and embryo quality, this retrospective study analyzed 151 intracytoplasmic sperm injection (ICSI) cycles (1152 embryos collected) between November 2016 and June 2019. SDF was assessed using sperm chromatin dispersion. The cycles were divided into two groups based on the SDF rate: SDF < 15% (n = 114) and SDF ≥ 15% (n = 37). The embryo morphokinetic parameters, cleavage patterns, and embryo quality were compared between the two groups. The morphokinetic parameters tPNf, t2, t3, t4, t5, t6, and t8 were achieved significantly earlier in the SDF < 15% group compared with in the SDF ≥ 15% group. The fertilization and 2PN rates seemed to be significantly higher in the SDF < 15% group compared with in the SDF ≥ 15% group, while the abnormal cleavage rates were similar. However, a significantly higher rate of chaotic cleavage (CC) was observed in the SDF ≥ 15% group. The D3 high-quality embryo and available embryo rates were similar between the two groups. The blastocyst formation, high-quality blastocyst, and available blastocyst rates in the SDF < 15% group were significantly higher than those in the SDF ≥ 15% group. With an increase in SDF level, the chemical pregnancy, clinical pregnancy and implantation rates tended to decrease, while the miscarriage rate increased. This study demonstrated that SDF ≥ 15% reduces the fertilization rate of ICSI cycles and affects certain morphokinetic parameters. A higher SDF level can also induce a higher rate of CC, with subsequent decreases in the blastocyst formation rate and blastocyst quality.

The effects of Pb on sperm parameters and sperm DNA fragmentation of men investigated for infertility

2020 ◽  
Vol 31 (4) ◽  
Author(s):  
G.U.S. Wijesekara ◽  
D.M.S. Fernando ◽  
S. Wijeratne
Keyword(s):  
Dna Fragmentation ◽  
Seminal Plasma ◽  
Sperm Concentration ◽  
Sperm Parameters ◽  
World Health ◽  
Sperm Dna Fragmentation ◽  
Progressive Motility ◽  
Sperm Dna ◽  
The Mean ◽  
Pb Concentration

AbstractBackgroundLead (Pb) is one of the metals most prevalent in the environment and is known to cause infertility and deoxyribonucleic acid (DNA) fragmentation. This study aimed to determine the association between seminal plasma Pb and sperm DNA fragmentation in men investigated for infertility.MethodsMale partners (n = 300) of couples investigated for infertility were recruited after informed consent was obtained. Sperm parameters were assessed according to the World Health Organization (WHO) guidelines. Seminal plasma Pb was estimated by atomic absorption spectrophotometry after digestion with nitric acid.ResultsIn Pb-positive and -negative groups the sperm parameters and sperm DNA fragmentation were compared using independent sample t-test and the Mann-Whitney U-test, respectively. The mean [standard deviation (SD)] age and duration of infertility were 34.8 (5.34) years and 45.7 (35.09) months, respectively, and the mean Pb concentration was 15.7 μg/dL. In Pb positives compared to Pb negatives the means (SD) of sperm count, progressive motility viability and normal morphology were lower (p > 0.05) but the DNA fragmentation was significantly higher 39.80% (25.08) than Pb negatives 22.65% (11.30). Seminal plasma Pb concentration and sperm DNA fragmentation had a positive correlation (r = 0.38, p = 0.03). A negative correlation was observed between sperm DNA fragmentation and sperm concentration, progressive motility, total motility and viability. When the DNA fragmentation was ≥30% sperm concentration and viability decreased (p < 0.05).ConclusionsPb in seminal plasma had a significant effect on sperm DNA fragmentation but not with other sperm parameters.

scholarly journals 272 Proposing a combination of heritable fertility traits for bull selection

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 83-84
Author(s):  
Marina Fortes ◽  
Wei Liang Andre Tan ◽  
Laercio R Porto-Neto ◽  
Antonio Reverter ◽  
Gry B Boe-Hansen
Keyword(s):  
Dna Fragmentation ◽  
X Chromosome ◽  
Selective Breeding ◽  
Genetic Correlations ◽  
Research Data ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Fertility Traits ◽  
Protamine Deficiency

Abstract Traits such as sperm morphology and motility are routine in veterinarian evaluations of bull fertility. However, they rarely are included in livestock breeding programs, which typically use only scrotal circumference (SC) and some female traits for fertility selection. We studied 25 male fertility traits measured in two research populations of bulls (1,099 Brahman, and 1,719 Tropical Composite) and one commercial population (2,490 Santa Gertrude bulls). Measurements included standard semen evaluation (e.g. sperm motility and morphology) and SC. In the research data, we also measured sperm DNA fragmentation and sperm protamine deficiency for about 50% of the bulls. Using a mixture of genomic and pedigree analyses, we estimated heritabilities and genetic correlations for all traits, in each population. Our analyses suggest that bull fertility traits have a heritable component, which makes selective breeding possible. The phenotype variation in sperm DNA fragmentation and sperm protamine deficiency traits also have a heritable component (h2 ~ 0.05–0.22). These first estimates for heritability of sperm chromatin phenotypes require further studies, with larger datasets, to corroborate present results. In all three populations, we observed genetic correlations across traits that were favorable, but not high. For example, the percentage of normal sperm (PNS) from the sperm morphology evaluation was positively correlated with SC. In the research data, sperm DNA fragmentation was negatively correlated with PNS (r2 ~ 0.23–0.33), meaning that bulls with a higher PNS had less DNA fragmentation, being therefore more fertile according to both indicators. Given the favorable and yet not high genetic correlations between traits, it is possible to envision that sperm chromatin phenotypes might form a panel, together with PNS and SC, for a comprehensive bull fertility index. Selection indices that include fertility traits are being implemented in the dairy industry and could be recommended for beef cattle, too. An index that benefits from the favorable genetic correlations between traits that describe different aspects of bull fertility is a sensible approach to selective breeding. The clinical use of complementary indicators for male fertility is largely accepted, when deciding on bull fitness for the mating season. We propose extending this rationale to create a multi-trait index that captures genetic merit for bull fertility. In addition, we performed genome-wide association analyses in the research data and identified eight QTLs in the X chromosome. Correlations and shared SNP associations support the hypothesis that these phenotypes have the same underlying cause: abnormal spermatogenesis. In conclusion, it is possible to improve bull fertility through selective breeding, by measuring complementary fertility traits. Genomic selection for bull fertility might be more accurate if the X chromosome mutations that underlie the discovered QTL are included in the analyses. Polymorphisms associated with fertility in the bull accumulate in the X chromosome, as they do in humans and mice, thus suggesting specialization of this chromosome.

The assessment of sperm DNA fragmentation in the saltwater crocodile (Crocodylus porosus)

2017 ◽  
Vol 29 (3) ◽  
pp. 630 ◽  
Author(s):  
S. D. Johnston ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
J. L. Fernández ◽  
J. Gosálvez
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmented Dna ◽  
Sperm Dna ◽  
Saltwater Crocodile ◽  
Crocodylus Porosus ◽  
Dispersion Test ◽  
Sperm Nuclei

Herein we report a method of assessing DNA fragmentation in the saltwater crocodile using the sperm chromatin dispersion test (SCDt) after including frozen–thawed spermatozoa in a microgel (Halomax; Halotech DNA, Madrid, Spain). Following controlled protein depletion, which included a reducing agent, sperm nuclei with fragmented DNA showed a homogeneous and larger halo of chromatin dispersion with a corresponding reduced nucleoid core compared with sperm with non-fragmented DNA. The presence of DNA damage was confirmed directly by incorporation of modified nucleotides using in situ nick translation (ISNT) and indirectly by studying the correlation of the SCDt with the results of DNA damage visualisation using a two-tailed comet assay (r = 0.90; P = 0.037). Results of the SCDt immediately following thawing and after 5 h incubation at 37°C in order to induce a range of DNA damage revealed individual crocodile differences in both the baseline level of DNA damage and DNA longevity.

Evaluating human sperm DNA integrity: relationship between 8-hydroxydeoxyguanosine quantification and the sperm chromatin structure assay

Zygote ◽  
2003 ◽  
Vol 11 (4) ◽  
pp. 367-371 ◽  
Author(s):  
Isabelle Oger ◽  
Christelle Da Cruz ◽  
Gilles Panteix ◽  
Yves Menezo
Keyword(s):  
Dna Fragmentation ◽  
Chromatin Structure ◽  
Inverse Relationship ◽  
Sperm Concentration ◽  
Sperm Chromatin ◽  
Sperm Chromatin Structure Assay ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Oxidative Products ◽  
Dna Fragmentation Index

In our work, we have used 8-hydroxy-deoxyguanosine (8-OH-dG), one of the major oxidative products of sperm DNA, in a population of patients consulting for infertility. We found an inverse relationship between sperm concentration and the log of the ratio of 8-OH-dG to dG (P<0.01). On the same patients' sperm samples, the sperm chromatin structure assay (SCSA) was performed. An inverse relationship was observed between the DNA fragmentation index and sperm concentration (P<0.001). There was also a positive relationship between SCSA and log 8-OH-dG/dG. This indicates that DNA fragmentation measured by the SCSA originates in part from oxidative products. In a few patients, antioxidant treatment decreased the DNA fragmentation index below the threshold of 30% that is crucial for subfertility.

scholarly journals Sperm DNA fragmentation in Italian couples with recurrent pregnancy loss

2017 ◽  
Vol 34 (1) ◽  
pp. 58-65 ◽  
Author(s):  
Tania Carlini ◽  
Donatella Paoli ◽  
Marianna Pelloni ◽  
Fabiana Faja ◽  
Alessandro Dal Lago ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Pregnancy Loss ◽  
Recurrent Pregnancy Loss ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna
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