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.

Influence of obesity on sperm DNA fragmentation index and outcomes of IVF programs

2015 ◽  
Vol 61 (5) ◽  
pp. 48-55 ◽  
Author(s):  
Irina Ivanovna Vitiazeva ◽  
Marina Victorovna Altashina ◽  
Tatiana Vladimirovna Mun ◽  
Ekaterina Anatolievna Troshina
Keyword(s):  
Body Mass Index ◽  
Dna Fragmentation ◽  
Body Mass ◽  
Reproductive Age ◽  
Semen Parameters ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

The reduction of the birth rates in the developed countries and increase in the frequency of male infertility stimulate the extensive investigations for the factors that negatively affect the reproductive system of the men and causing their infertility. The excessive body weight and obesity in the men of the reproductive age can promote the development of infertility. One of the mechanisms by which excess fat tissue has a negative impact on male fertility is disturbance of spermatogenesis. The authors aggregate scientific publications concerning the macroscopic and ultrastructural disturbances of spermatogenesis in men with obesity. We present the results of the study conducted at the Department of ART Endocrinology Research Center, targeted at the revelation of the relationship of body mass index of men of reproductive age, semen parameters, sperm DNA fragmentation index, as well as the influence of body mass index on outcomes of in vitro fertilization programs.

82 DNA FRAGMENTATION DYNAMICS OF KOALA SPERMATOZOA

2009 ◽  
Vol 21 (1) ◽  
pp. 141 ◽  
Author(s):  
Y. P. Zee ◽  
C. Lopez-Fernandez ◽  
J. Gosalvez ◽  
W. V. Holt ◽  
S. D. Johnston
Keyword(s):  
Dna Fragmentation ◽  
Dna Degradation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Rate Of Increase ◽  
Fragmentation Dynamics ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

Koala sperm chromatin has a tendency to relax following incubation and thawing but the background incidence and dynamics of DNA fragmentation during semen processing at 35°C and following chilled and frozen preservation has not been investigated. This study (n = 10) was designed to establish the fragmentation dynamics of koala sperm DNA at body temperature (35°C), after chilling (4°C) for upward of 16 days, and following a standard freeze–thaw protocol (Johnston SD et al. 2006 Cryobiology 53, 218–228). Sperm DNA fragmentation index (sDFI) was determined using a Halomax kit (ChromaCell SL, Madrid, Spain), which had been customized and validated for koalas (Johnston SD et al. 2007 J. Androl. 28, 891–899). All semen was assessed for sDFI over a 48-h incubation period (T0, T2, T6, T24, and T48) at 35°C. After incubation at 35°C for 48 h, the sDFI and rate of DNA degradation of freshly diluted spermatozoa were highly variable between individuals; the sDFI for 2 koalas remained consistently low (≤2%) whereas the other 8 had sDFI of 8 to 12% after incubation. Chilled storage increased sDFI in all animals, but the rate of increase and the time at which the DNA started to fragment also varied between koalas; sDFI for 1 koala increased immediately upon rewarming after being chilled for 4 h, whereas that of another koala did not increase until after 8 days of chilling and 24 h of incubation at 35°C. Animals also responded to cryopreservation differently; sDFI increased after thawing for 2 of the koalas but did not increase in the others. Subsequent evaluation of frozen–thawed spermatozoa from a greater number of captive koalas (n = 22), and under extended conditions of post-thaw incubation (up to 17 days at 35°C) permitted categorization of the koalas into 3 distinctive groups based on their DNA fragmentation dynamics and rate of DNA degradation. For 7 of the animals, sDFI remained close to the basal level when incubated at 35°C over 7 days, whereas 2 of the koalas had sDFI ranging from 40 to 70% after 24 h of incubation. This study confirmed the occurrence of inter-animal variability in the dynamics of DNA fragmentation, a finding that was apparent whether or not the spermatozoa had been subjected to chilling or cryopreservation.

scholarly journals Correlation between sperm DNA fragmentation index and semen parameters in 418 men seen at a fertility center

2021 ◽  
Author(s):  
Láyonal Germán Acosta Campos ◽  
Lissett Chiscul Requejo ◽  
Carlos Antonio Rivas Miñano ◽  
Jheny Díaz Orrego ◽  
Elmer Chávez Loyaga ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Semen Parameters ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

84 EFFECT OF A STRESSOR ON CANINE SPERM DNA FRAGMENTATION USING THE SPERM CHROMATIN DISPERSION TEST

2013 ◽  
Vol 25 (1) ◽  
pp. 189
Author(s):  
M. Urbano ◽  
J. Dorado ◽  
I. Ortiz ◽  
M. J. Galvez ◽  
S. Demyda-Peyras ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Dispersion Test ◽  
Fresh Semen ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index ◽  
Sperm Chromatin Dispersion Test

Recently, a new procedure for the analysis of sperm DNA fragmentation has been developed for human and different mammalian species (Sperm-Halomax®), based on the sperm chromatin dispersion test (SCDt); however, no studies has been performed specifically on canine frozen–thawed-stressed semen but is there for cooled semen. The aim of this work was to assess the effect of a stressor (24 h in an oven at 38°C) on canine frozen–thawed semen using the SCDt to resemble what happens in the female reproductive tract. For this purpose, ejaculates were collected by digital manipulation from 4 healthy beagle dogs and the sperm-rich fraction of the ejaculates from 3 different dogs was pooled each time. All the pooled semen samples (n = 4) used presented physiological values concerning to routine semen parameters (motility, morphology, and sperm concentration). After evaluation, semen samples were centrifuged and the sperm pellet resuspended to a final concentration of 100 × 106 sperm mL–1 in 2 steps with CaniPRO Freeze (Minitub, Tiefenbach, Germany). Sperm were slowly cooled to 5°C and then loaded into 0.5-mL plastic straws. After that, straws were frozen in liquid-nitrogen vapours for 10 min and stored into a nitrogen tank. Straws were thawed in a water bath (30 s/37°C) and incubated for 24 hours at 38°C before analysis. The sperm DNA fragmentation was assessed in fresh semen and frozen–thawed-stressed samples using the Sperm-Halomax® commercial kit specifically developed for canine semen (Halotech DNA SL, Madrid, Spain) following the manufacturer’s instructions. Slides were stained for green fluorescence staining and 500 sperm per slide were counted using fluorescence microscopy. The sperm DNA fragmentation index (%) was compared between fresh and frozen–thawed-stressed semen samples by ANOVA. Results were expressed as mean ± standard error of the mean. The results obtained showed that subjecting thawed semen to 24 h in an oven at 38°C significantly increased (P < 0.05) DNA fragmentation compared with fresh semen (2.7% ± 0.2 v. 1.4 ± 0.1%). The stress factor was performed to simulate the viability of canine thawed sperm (12–24 h) when a bitch is inseminated with frozen semen. It would be interesting to perform further studies to relate sperm DNA fragmentation and fertility of frozen–thawed canine semen. In conclusion, frozen–thawed-stressed semen samples increased the sperm DNA fragmentation index measured using a SCDt. Further studies are needed to relate sperm DNA fragmentation with fertility rates or cryopreservation success.

scholarly journals Microscopic Varicocelectomy Significantly Decreases the Sperm DNA Fragmentation Index in Patients with Infertility

2014 ◽  
Vol 2014 ◽  
pp. 1-4 ◽  
Author(s):  
Teoman Cem Kadioglu ◽  
Emin Aliyev ◽  
Murad Celtik
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Infertile Men ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

Background. Varicocele is associated with high levels of DNA damage in spermatozoa due to oxidative stress and elevated levels of sperm DNA fragmentation, which has been currently proposed to be an essential additional diagnostic test to be recommended for patients with clinical varicocele. The aim of this study was to evaluate the parameters of semen and the DNA fragmentation index (DFI) in patients with varicocele before and after varicocelectomy.Methods. The details of 92 consecutive patients were retrospectively analyzed from January 2010 to December 2012. The sperm samples were evaluated according to the World Health Organization Guidelines. Sperm DNA damage, characterized as DFI, was evaluated by sperm chromatin structure assay using flow cytometry.Results. There was a statistically significant improvement in the semen concentration, the total motile count, the total normal sperm count, and the sperm DNA fragmentation index (DFI; the percentage of sperm with denatured DNA) after varicocelectomy. There was a large decrease in DFI from a preoperative mean of 42.6% to a postoperative mean of 20.5% (P<0.001). A higher preoperative DFI was associated with a larger decrease in postoperative DFI, and significant negative correlations were observed between the DFI and sperm motility (r=-0.42,P<0.01).Conclusion. Our data suggest that varicocelectomy can improve multiple semen parameters and sperm DNA damage in infertile men with varicocele. The patients with preoperative defects in those parameters showed greater improvement postoperatively. Further research in this area is needed to understand the exact mechanisms of DNA damage in infertile men with varicocele.

scholarly journals Correlation between Sperm Dna Fragmentation and Conventional Semen Parameters among Different Age Groups

2021 ◽  
Vol 14 (3) ◽  
pp. 1345-1350
Author(s):  
Shruti Chopra ◽  
Ajit Varma ◽  
Seema Jain ◽  
Sangeeta Jain ◽  
Devendra Choudhary
Keyword(s):  
Dna Fragmentation ◽  
Staining Method ◽  
Age Groups ◽  
Sperm Concentration ◽  
Semen Parameters ◽  
Sperm Dna Fragmentation ◽  
Normal Morphology ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Dna Fragmentation Index

Objective: To study the relationship between conventional semen parameters and sperm chromatin condensation (DNA fragmentation index) using aniline blue-eosin staining method among patients of different age groups visiting the In-vitro fertilization (IVF) clinic.Design: Retrospective study Setting: Tertiary care infertility centre Method: A total of 240 patient semen samples were studied between the period of May 2015 to May 2016 for conventional semen parameters (WHO criteria) and DNA fragmentation index (DFI) using aniline blue- eosin staining method. Patients were separated into three groups: <=30 years, 31-35 years and 36 years & above. Statistical analysis was performed using Pearson correlation co-efficient and regression tests on the groups. Main Outcome Measures: Sperm concentration (Millions /ml), motility(%), normal morphology(%), DFI (%). Result: In each age group, i.e., <=30years, 31-35 years and 36 years & above, there was a significant and negative correlation between DFI and sperm concentration (r= -0.50, r= -0.34, r= -0.49 respectively; P<0.05), motility(r= -0.69,r= -0.66, r= -0.54 respectively; P<0.05) and normal morphology (r= -0.86,r= -0.80, r= -0.75 respectively; P<0.05). Sperm DNA fragmentation index among the age groups was not statistically significantly (P>0.05). Conclusion: Our study demonstrated that age is not a predictor of DFI. Whereas, sperm concentration, sperm motility and normal sperm morphology showed a significant association with DFI in all the age groups i.e., better the conventional semen parameters, lower the DFI.

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