scholarly journals The risk of sperm demage in men with the combined effect of endocrine disruptors

2019 ◽  
Vol 74 (4) ◽  
pp. 229-234
Author(s):  
Stanislav V. Chigrinets ◽  
Gennadiy V. Brukhin
Keyword(s):  
Dna Damage ◽  
Bisphenol A ◽  
Dna Fragmentation ◽  
Endocrine Disruptors ◽  
Seminal Fluid ◽  
Combined Effect ◽  
Sperm Cells ◽  
Median Concentration ◽  
Sperm Dna ◽  
Comparison Groups

Background: The most frequent ultrastructural change in sperm cells is nuclear DNA fragmentation. Many authors believe that DNA damage to sperm cells can serve as a diagnostic marker of a negative paternal effect in the pre-implantation period of human development. A number of studies have shown a direct correlation between the high percentage of sperm DNA damage and the frequency of spontaneous abortions. On the other hand, it is known that the damaging effect of endocrine disruptors, for example, triclosan and bisphenol A, is based on their ability to induce oxidative stress, which is considered as one of the factors in cell DNA damage. Aims: to assess the risk of sperm DNA fragmentation in men with the combined effect of bisphenol A, triclosan and 4-nonylphenol in seminal fluid. Materials and methods: 84 samples of seminal fluid of men with normo-and patozoospermia were studied. The concentration of bisphenol A, triclosan and 4-nonylphenol in gas was determined by gas chromatography with mass spectrometry (GC-MS). The comparison groups were divided according to the degree of DNA fragmentation of spermatozoa into two groups: the 1st group of patients with DNA fragmentation of spermatozoa 15% (n=18) and the 2nd group 15% (n=29). The spermiological study was carried out according to the WHO recommendations (2010) taking into account the assessment of the number of spermatozoa, their motility and morphology, as well as the degree of fragmentation of the sperm DNA. Results: Bisphenol A was found in 100% of the ejaculate samples with a median concentration of 0.150 ng/ml. Triclosan and 4-nonylphenol were detected in 84.3 and 98.1% of ejaculate samples with a median concentration of 0.11 and 0.16 ng/ml respectively. Comparison groups were statistically significantly distinguished by the concentration of bisphenol A and triclosan p=0.014; p0.001 respectively. Triclosan with an increase in concentration in seminal fluid by 0.1 ng/ml increased the chance of development of the degree of DNA fragmentation 15% by 2.9 times. The odds ratio of bisphenol A and 4-nonylphenol to DNA damage was not statistically significant. The prognostic model of the joint effect of endocrine disruptors on DNA damage, constructed using multiple logistic regression analysis, was also found to be statistically significant only with respect to the effect of triclosan. Conclusions: The risk of damage to sperm DNA in men is primarily associated with the effect of triclosan in seminal fluid. At the same time, it is necessary to assume the absence of a synergistic effect of bisphenol A, triclosan and 4-nonylphenol on the DNA damage of spermatozoa in men.

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.

scholarly journals Changes in the Level of DNA Fragmentation in Sperm Cells detected by Acridine Orange Test in Men with Sub/infertility Treated with Nutritional Supplement PAPA

Folia Medica ◽  
2020 ◽  
Vol 62 (1) ◽  
pp. 112-116
Author(s):  
Stoil N. Tomov ◽  
Radoslava St. Stoyanova ◽  
Pepa K. Atanasova ◽  
Ivan Y. Dechev
Keyword(s):  
Acridine Orange ◽  
Dna Fragmentation ◽  
Physiological Condition ◽  
Nutritional Supplement ◽  
Sperm Cells ◽  
Sperm Dna Fragmentation ◽  
Double Stranded Dna ◽  
Sperm Dna ◽  
Before And After ◽  
Sperm Nuclei

Background: When diagnosing and treating male infertility it is important to determine whether there are defects in the maturation process of sperm nuclei. Using nutritional supplements can improve the morphological and physiological condition of the spermatozoa. In recent years there has been an increase in the usage of supplements with different compositions which strives to determine the best combination and avoid side effects. &nbsp; Aim: To study the effect of PAPA nutritional supplement on the levels of DNA fragmentation of sperm cells tested with acridine orange test (single stranded DNA against double stranded DNA) in men with sub/infertility. &nbsp; Materials and methods: 48 men with confirmed sub/infertility underwent treatment for three months with nutritional supplement PAPA containing 9 micronutrients. The differences in levels of DNA fragmentation were determined with acridine orange test, which was conducted before and after the treatment. &nbsp; Results: The results were statistically significant (p<0.001) showing an increase in the number of green spermatozoa (normal DNA), and a decrease of damaged ones (orange and red). After treatment the level of sperm DNA fragmentation decreased by 10.2%. &nbsp; Conclusion: Men with confirmed sub/infertility that took nutritional supplement PAPA for three moths showed a decrease in DNA fragmentation levels of 10.2% determined by AO test which implies an improvement of male fertility levels.

P–006 Simultaneous determination of bisphenol A and S in the samples of human seminal fluid

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Fialková ◽  
T Král ◽  
J Kohoutek ◽  
K Franzová ◽  
M Ješeta ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Bisphenol A ◽  
Endocrine Disruptors ◽  
Analytical Method ◽  
Seminal Fluid ◽  
Limit Of Detection ◽  
Semen Parameters ◽  
Ms Detection ◽  
Validation Criteria ◽  
Limit Of Quantitation

Abstract Study question Can we quantitatively determine concentrations of endocrine disruptors namely bisphenol A and S in seminal fluid? Summary answer We developed selective analytical method to simultaneously screen for the presence of bisphenol A (BPA) and S (BPS). What is known already The male reproductive system involves processes, which may be influenced by the disruption of the endocrine system by chemicals called endocrine disruptors (EDs). There is a growing evidence that EDs such as bisphenol A and S may be responsible for the decline in male reproductive health. To date, the claimed adverse effects on male fertility are largely based on the results from studies assessing the relationship between urinary BPA and BPS concentration and semen parameters. The best evidence of an adverse effect of BPA and BPS directly on spermatozoa could be provided by measuring bisphenols concentration directly in seminal fluid. Study design, size, duration To selectively and quantitatively analyzed bisphenols in any biological matrix advanced analytical tools and selective sample preparation protocols must be employed. In this study we developed targeted analytical method based on liquid chromatography tandem mass (LC-MS/MS) detection to measure bisphenol A and S in seminal fluid samples obtained from IVF clinic. A total of 140 samples were analysed. Participants/materials, setting, methods BPA and BPS was extracted from 140 seminal fluid samples using solvent extraction followed by preconcentration step. Samples were analyzed on Agilent 6495 Triple Quadrupole (Agilent Technologies, Santa Clara, CA) operating in the ESI-negative mode. Two MS/MS transitions were used for quantitative LC-MS/MS analyses. Chromatographic separation was achieved on Waters™ ACQUITY™ UPLC™BEH C18 (100 × 2.1 mm, 1.7 µm) column using gradient elution with a mixture of 0.1mM ammonium fluoride and methanol as mobile phases. Main results and the role of chance We developed selective sample preparation method for detection of BPA and BPS in seminal fluid followed by LC-MS/MS detection. The method validation was performed based on FDA guidelines. Validation criteria included limit of detection (LOD), limit of quantitation (LOQ), accuracy and precision. Due to the lack of the certified reference material the validation criteria of the method were assessed in pool of spiked seminal samples. The accuracy of the LC-MS/MS method was evaluated as a percent recovery of the amount of target analyte added into the sample. Recovery rates were above 80% for both analytes. LOD was 0.04 ng/mL for BPA and 0.01 ng/mL for BPS. LOQ was 0.14 ng/mL and 0.02 ng/mL for BPS. Measured BPA concentration ranged from 0.04 ng/mL to 1.62 ng/mL. For BPS, the concentration ranged from 0.01 ng/mL to 0.47 ng/mL. BPA and BPS were detected in 64% and 81% of samples, respectively. Interestingly, BPA showed lower detection frequency compared to BPS. These results are consistent with other studies performed on urine samples. Limitations, reasons for caution The limitation of the developed method is the time-consuming sample preparation and analysis cost. Wider implications of the findings: These results document for the first time the presence of BPS in seminal fluid. Knowing the concentration of BPA and BPS in seminal fluid is crucial for mitigating the associated health risks and initiating intervention and prevention strategies. Our future work will evaluate the influence of BPS concentration on spermatozoa. Trial registration number AZV NV18–01–00544; CZ.02.2.69/0.0/0.0/19_074/0012727

Evaluation of Sperm DNA Fragmentation via Halosperm Technique and TUNEL Assay Before and After Cryopreservation

2019 ◽  
Vol 26 (12) ◽  
pp. 1575-1581 ◽  
Author(s):  
Senay Cankut ◽  
Turgay Dinc ◽  
Mehmet Cincik ◽  
Guler Ozturk ◽  
Belgin Selam
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Tunel Assay ◽  
University Hospital ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sperm Dna Fragmentation ◽  
Liquid Form ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Before And After

Aim: Human sperm DNA fragmentation is one of the factors suggested for male infertility. The ratio of sperm DNA damage in semen may adversely affect both the fertilization rate and the embryo development of in vitro fertilization/ intracytoplasmic sperm injection cycles. Sperm cryopreservation both increases the success rates in assisted reproductive techniques (ARTs) and contributes to the preservation of fertility before testis surgery, chemotherapy, and radiotherapy. The aim of the current study is to determine sperm DNA fragmentation, following cryopreservation. Methods: A cross-sectional, observational study was conducted at a university hospital infertility clinic. One hundred (n = 100) volunteer fertile men (ages between 21 and 39 years) with normozoospermic sperm parameters were involved in the current study. Sperm DNA damage was evaluated with the Halosperm technique and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Fresh samples were studied in liquid form. The remaining samples were kept frozen and then thawed after 1 month and reevaluated with the Halosperm technique and TUNEL assay. Results were then compared between the fresh and frozen samples. Results: Sperm DNA fragmentation results with the Halosperm technique both before and after cryopreservation were 25% (5%-65%) and 40% (6%-89%), respectively, with a statistically significant increase (15%; P < .001). Sperm DNA fragmentation results by TUNEL assay before and after cryopreservation were 17% (3%-43%) and 36% (7%-94%), respectively, with a statistically significant increase (19%; P <.001). Conclusion: The current data demonstrate increased sperm DNA damage after cryopreservation. Further studies may contribute to development of less harmful techniques and cryoprotectants in order to improve the results of ART.

scholarly journals Sperm DNA Fragmentation: Origin and Impact on Human Reproduction

2011 ◽  
Vol 2 (2) ◽  
pp. 88-108 ◽  
Author(s):  
Ralf R. Henkel ◽  
Daniel R. Franken
Keyword(s):  
Dna Damage ◽  
Environmental Factors ◽  
Dna Fragmentation ◽  
Human Reproduction ◽  
Sperm Dna Fragmentation ◽  
Permanent Damage ◽  
Sperm Dna Damage ◽  
Male Genome ◽  
Sperm Dna ◽  
Early Abortion

Sperm DNA can be damaged due to a multitude of different noxae, which include disease, and occupational and environmental factors. Depending on the magnitude of the damage, such lesions may be repaired by the oocyte or the embryo. If this is not possible, a permanent damage can be manifested leading to mutations of the male genome. In cases where the oocyte or the embryo does not counter these damages to the male genome in terms of repair or an early abortion, sperm DNA damage and fragmentation can be a cause of numerous diseases including childhood cancer.

scholarly journals Rapid rates of sperm DNA damage after activation in tench (Tinca tinca: Teleostei, Cyprinidae) measured using a sperm chromatin dispersion test

Reproduction ◽  
2009 ◽  
Vol 138 (2) ◽  
pp. 257-266 ◽  
Author(s):  
Carmen López-Fernández ◽  
Matthew J G Gage ◽  
Francisca Arroyo ◽  
Altea Gosálbez ◽  
Ana M Larrán ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Dna Fragmentation ◽  
Living Organism ◽  
Sperm Chromatin ◽  
Tinca Tinca ◽  
External Fertilization ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Dispersion Test

Spermatozoal haplotypic DNA is prone to damage, leading to male fertility problems. So far, the assessment of sperm DNA breakage has been challenging because protamines render the nuclear chromatin highly compacted. Here, we report the application of a new test to quantify DNA fragmentation in spermatozoa of an externally fertilizing teleost fish. The sperm chromatin dispersion (SCD) test uses a species-specific lysing solution to generate controlled protein depletion that, followed by DNA-specific fluorescent labelling, allows an easy morphological discrimination between nuclei affected by DNA damage. Using tench (Tinca tinca) as our model, we first trialled the test against established, but more technically demanding, assays employing in situ nick translation (ISNT) and the comet assay. The SCD test showed high concordance with ISNT, comet assay measures and a chromatin-swelling test, confirming the application of this straightforward SCD technique to various aspects of reproductive biology. Second, we examined between-male variation in DNA damage, and measured changes through time following spermatozoal activation. Between-male variation in the basal levels of average DNA damage ranged from 0 to 20% of sperm showing damage, and all showed increases in DNA fragmentation through time (0–60 min). The rates of DNA damage increase are the fastest so far recorded in sperm for a living organism, and may relate to the external fertilization mode. Our findings have relevance for broodstock selection and optimizing IVF protocols routinely used in modern aquaculture.

scholarly journals Effect of sperm DNA fragmentation on ICSI outcome: A prospective study

2020 ◽  
Vol 3 (2) ◽  
pp. 127-131
Author(s):  
Lakshamanan Saravanan ◽  
Mahalakshmi Saravanan ◽  
Ramya Harish ◽  
Nidhi Sharma
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
No Reduction ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Secondary Objective ◽  
A Prospective Study ◽  
Thesis Work ◽  
Embryo Grade

Aim and objectives: The primary aim was to measure the sperm DNA damage and to study the magnitude of sperm DNA damage. Secondary objective was to study the effect of sperm DNA fragmentation on Day 5 Blastocyst expansion (graded 1-5). Results: There is an increase in sperm DNA fragmentation with an increase in age. Increased sperm DNA fragmentation is also associated with abnormal motility and morphology in semen samples. However, there is no reduction in expansion or grade of blastocyst. Conclusion: Sperm DNA fragmentation testing is a useful investigation in unexplained infertility. However, Sperm DNA fragmentation has no significant association with Day 5 embryo grade in ICSI cycles. Thesis work of Fellowship in Reproductive Medicine student: Dr. Ramya Harish

283 DNA FRAGMENTATION DYNAMICS AND POST-THAW MOTILITY OF WHITE-TAILED DEER SPERM

2011 ◽  
Vol 23 (1) ◽  
pp. 239 ◽  
Author(s):  
M. E. Kjelland ◽  
C. González-Marín ◽  
J. Gosálvez ◽  
C. López-Fernández ◽  
R. W. Lenz ◽  
...  
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Technical Assistance ◽  
Wilcoxon Test ◽  
Sperm Dna Fragmentation ◽  
Kaplan Meier ◽  
Sperm Dna ◽  
Fort Collins ◽  
Live Sperm ◽  
Conservation Of Endangered Species

The main objective herein was to study the level of DNA damage and post-thaw motility of White-tailed deer sperm before (neat sample) and after sex-sorting and conventional-sorting using a MoFlo® SX flow cytometer (SX, Dako, Fort Collins, CO, USA). For assessing DNA damage, a comparison of frozen–thawed (F-T) neat sperm (control) was made with F-T sex-sorted, F-T conventional-sorted, and F-T conventional sperm samples. Sperm motility was assessed by bright field microscopy using a Nikon Eclipse 80i microscope and slide-coverslips (25.4 × 76.2 mm slides, 22 × 22 #1.5 coverslips). A direct comparison of all 4 aforementioned sperm groups could not be made for some bucks. Live/dead sorting of the sperm (i.e. conventional-sorted sperm) can remove membrane compromised sperm and nonaligned live sperm, which may result, in part, from abnormal morphologies. White-tailed deer (Odocoileus virginianus; n = 13) from 1 to 7 years old were used for the experiments. The White-tailed deer were selected based on a genetic predisposition for producing large antlers (i.e. Boone and Crockett antler scores ≥200 points). Sperm DNA fragmentation levels were assessed using the Sperm-Halomax® kit (Halotech DNA, Madrid, Spain), counting 300 sperm per sample. The level of baseline DNA fragmentation was similar for conventional F-T sperm samples (<5%), but even lower after sex-sorting and conventional-sorting (2.39 and 1.69%, respectively). The conventional sperm samples had lower post-thaw motilities compared with sex-sorted samples from the same individual bucks (n = 6), with average post-thaw motilities of 43 ± 26% and 56.5 ± 20%, respectively. The statistical comparison of the dynamic loss of DNA quality (i.e. DNA fragmentation of samples incubated in a 34°C water bath for 96 h) was assessed using the nonparametric maximum likelihood Kaplan-Meier estimator and a Breslow (Generalized Wilcoxon) test. When comparing sperm samples taken from the same bucks (n = 6), the conventional samples had significantly greater (P < 0.05) DNA fragmentation levels over time than the sex-sorted sperm. Conventional-sorted White-tailed deer (n = 8) sperm samples did not have significantly greater (P > 0.05) DNA fragmentation levels when compared with the sex–sorted sperm. When comparing X-chromosome sorted sperm to Y-chromosome sorted sperm, the DNA fragmentation levels were not significantly different (n = 10; P > 0.05), averaging 2.59 ± 3.61% and 2.18 ± 0.53% after 96 h. Based on the sperm DNA fragmentation and post-thaw motility results in the present study, the sex-sorting of White-tailed deer sperm may be a viable technique for the White-tailed deer industry and perhaps serve as a model for the conservation of endangered species such as the Eld’s deer (Cervus eldithamin). Future work should be implemented for examining the fertilizing potential of sex-sorted White-tailed deer sperm. The authors thank Maurice Rosenstein, Laura Belluzzo, Jared Templeton, Mike Bringans, Pat Cooper, Suzanne Menges, Miguel Ramirez, Altea Gosálbez, and Sexing Technologies staff for technical assistance. This research was funded by Sexing Technologies.

scholarly journals Effect of bacterial infection on sperm quality and DNA fragmentation in subfertile men with Leukocytospermia

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Fatemeh Eini ◽  
Maryam Azizi Kutenaei ◽  
Fayegheh Zareei ◽  
Zeinolabedin Sharifian Dastjerdi ◽  
Maryam Hosseinzadeh Shirzeyli ◽  
...  
Keyword(s):  
Bacterial Infection ◽  
Dna Fragmentation ◽  
Bacterial Infections ◽  
Pathogenic Bacteria ◽  
Seminal Fluid ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

Abstract Background Although bacterial infections have been recognized as a possible cause of male infertility, the effect of bacterial infections on sperm quality and sperm DNA fragmentation remains controversial. The current study aimed to investigate the prevalence rate of bacterial infection in subfertile men and its effect on semen quality. Seminal fluid was collected from 172 male members of infertile couples attending the andrology infertility center and a group of 35 fertile subjects as a control. Sperm parameters and DNA fragmentation were evaluated based on the type of bacteria in all ejaculates. Results From the 172 patients investigated for infertility, 60 (34.88%) patients had a positive culture for pathogenic bacteria of different species. Leukocytospermia was significantly higher in infected samples in comparison with non-infected samples (p < 0.05). Sperm concentration and motility and morphology were significantly lower in infected than non-infected samples. Moreover, sperm DNA fragmentation was significantly higher in infected than non-infected samples. Besides, our results showed that sperm DNA fragmentation was correlated significantly with leukocytospermia (R: 0.22, p < 0.01). Conclusion The present study suggested that bacterial infection significantly correlated with leukocytospermia could impair male fertility potential through decreasing sperm motility, morphology, and DNA integrity.

scholarly journals Mitigating the Effects of Oxidative Sperm DNA Damage

Antioxidants ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 589
Author(s):  
Taylor Pini ◽  
Rachel Makloski ◽  
Karen Maruniak ◽  
William B. Schoolcraft ◽  
Mandy G. Katz-Jaffe
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Post Treatment ◽  
Semen Parameters ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Sperm Dna Fragmentation ◽  
Success Rates ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Cost Effective Treatment

Sperm DNA damage is correlated with reduced embryo development and increased miscarriage risk, reducing successful conception. Given its links with oxidative stress, antioxidants have been investigated as a potential treatment, yet results are conflicting. Importantly, individual antioxidants are not identical in composition, and some compounds may be more effective than others. We investigated the use of the polyphenol-rich, high-antioxidant-capacity fruit acai as a treatment for elevated sperm DNA fragmentation (>16%), measured by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). Following ≥ 74 days of treatment, we observed a significant decrease in sperm DNA fragmentation (−17.0% ± 2.5%) to 11.9 ± 1.7% (0–37%), with a 68.6% success rate (defined as post-treatment TUNEL < 16%). Post-treatment decreases in DNA fragmentation and success rates were not significantly impacted by low motility and/or concentration, or exceptionally high (> 25%) TUNEL. Treatment significantly reduced concentration in men with normal semen parameters, but 88% remained normal. Overall, successful treatment was not associated with age, semen parameters or TUNEL result at baseline. However, body mass index was significantly higher in nonresponders at baseline. This study provides evidence of a low-cost, effective treatment for elevated sperm DNA damage using acai.

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