scholarly journals A Modified Flotation Density Gradient Centrifugation Technique Improves the Semen Quality of Stallions with a High DNA Fragmentation Index

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1973
Author(s):  
Muhammad Umair ◽  
Heiko Henning ◽  
Tom A. E. Stout ◽  
Anthony Claes
Keyword(s):  
Dna Fragmentation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Quality Parameters ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Cushion Layer ◽  
Dna Fragmentation Index

Sperm DNA fragmentation compromises fertilization and early embryo development. Since spermatozoa lack the machinery to repair DNA damage, to improve the likelihood of establishing a healthy pregnancy, it is preferable to process ejaculates of stallions with a high sperm DNA fragmentation index (DFI) before artificial insemination or intracytoplasmic sperm injection. The aim of this study was to examine a modified flotation density gradient centrifugation (DGC) technique in which semen was diluted with a colloid solution (Opti-prepTM) to increase its density prior to layering between colloid layers of lower and higher density. The optimal Opti-prepTM solution (20–60%) for use as the bottom/cushion layer was first determined, followed by a comparison between a modified sedimentation DGC and the modified flotation DGC technique, using different Opti-prepTM solutions (20%, 25% and 30%) as the top layer. Finally, the most efficient DGC technique was selected to process ejaculates from Friesian stallions (n = 3) with high sperm DFI (>20%). The optimal Opti-prepTM solution for the cushion layer was 40%. The modified sedimentation technique resulted in two different sperm populations, whereas the modified flotation technique yielded three populations. Among the variants tested, the modified flotation DGC using 20% Opti-prepTM as the top layer yielded the best results; the average sperm recovery was 57%; the DFI decreased significantly (from 12% to 4%) and the other sperm quality parameters, including progressive and total motility, percentages of spermatozoa with normal morphology and viable spermatozoa with an intact acrosome, all increased (p < 0.05). In Friesian stallions with high sperm DFI, the modified flotation DGC markedly decreased the DFI (from 31% to 5%) and significantly improved the other semen quality parameters, although sperm recovery was low (approximately 20%). In conclusion, stallion sperm DFI and other sperm quality parameters can be markedly improved using a modified flotation DGC technique employing a 40% Opti-prepTM cushion and a 20% top layer.

scholarly journals Short communication. Stallion sperm quality after combined ejaculate fractionation and colloidal centrifugation

2015 ◽  
Vol 13 (4) ◽  
pp. e04SC02 ◽  
Author(s):  
Francisco Crespo ◽  
Jaime Gosálvez ◽  
Stephen D. Johnston ◽  
Joaquina De la Torre
Keyword(s):  
Dna Fragmentation ◽  
Sperm Quality ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Rate Of Increase ◽  
Sperm Sample ◽  
Artificial Vagina ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

<p>This study investigated the possible additive benefit of ejaculate fractionation and colloidal centrifugation on stallion sperm quality. Using an open-end artificial vagina, the sperm-rich fraction (FRAC-1) was separated from the rest of the ejaculate (FRAC-2) and a third sperm sample representing the combined ejaculate was reconstituted post-ejaculation (RAW). Each semen sample was processed for colloidal centrifugation. The percentage of abnormal spermatozoa was 17.8 ± 7.0% in RAW and 14.6 ± 9.5% in FRAC-1 but decreased to 11.4 ± 4.7% and 9.6 ± 6.9% respectively, after colloidal centrifugation. A sperm DNA fragmentation index of 10.9 ± 5.1% was observed in RAW and 7.5 ± 2.4% in FRAC-1 semen collected with the AV but this decreased to 7.8 ± 2.8% and 5.2 ± 2.3% after colloidal centrifugation. The rate of increase in sperm DNA fragmentation during the first 6 h of incubation at 37 ºC was 1.8 ± 0.9% per hour in RAW semen and 2.0 ± 2.0% per hour in FRAC-1 but this significantly decreased to 1.3 ± 1.4% and 0.9 ± 0.8% respectively after colloidal centrifugation. While stallion seminal characteristics can be improved using colloidal centrifugation, further enhancement is possible if the ejaculate is initially fractionated.</p>

DETERMINATION OF THE HUMAN SPERM DNA FRAGMENTATION INDEX

Vestnik ◽  
2021 ◽  
pp. 226-232
Author(s):  
К. К. Тлеуханов ◽  
Н. А. Алтыбаева ◽  
М. К. Отарбаев ◽  
Е. М. Тойшибеков ◽  
А. А. Тлеуханова
Keyword(s):  
Dna Fragmentation ◽  
Human Sperm ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

В статье представлены собранные данные о методах устранения повышенной частоты фрагментации ДНК у сперматозоидов, которые в некоторых исследованиях подтверждают, что введение антиоксидантов может снизить уровень фрагментации ДНК у сперматозоидов. The article presents collected data on methods of eliminating the increased frequency of DNA fragmentation in spermatozoa, which in some studies confirm that the introduction of antioxidants can reduce the level of DNA fragmentation in spermatozoa.

O-212 MACS vs TESA for raised sperm DNA fragmentation index – a RCT

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
K C Mantravadi ◽  
D R Gedela
Keyword(s):  
Dna Fragmentation ◽  
Live Birth ◽  
Trial Registration ◽  
Sperm Dna Fragmentation ◽  
Sperm Selection ◽  
Reproductive Outcomes ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Sperm Dna Fragmentation Index ◽  
Dna Fragmentation Index

Abstract Study question In Individuals with raised Sperm DNA Fragmentation Index (SDF), will sperm selection by magnetic activated cell sorting (MACS) or surgical retrieval of testicular sperms (TESA) optimize the reproductive outcomes? Summary answer Couples with failed implantation raised SDF, TESA /MACS offer similar results. This RCT doesn’t prove superiority or added benefit with any of the above interventions. What is known already It is evident that raised SDF negatively affects the reproductive outcomes. Management for raised SDF to optimize reproductive outcomes is still elusive. Study design, size, duration This was a Randomized Control Trial (RCT) with prior approval from institutional Ethical Committee and trial registration. Couples undergoing stimulation with raised SDF were randomized to MACS (n = 75) and TESA (n = 75) for sperm selection between April2019 & February2020. Participants/materials, setting, methods Couples with history of one failed IVF had SDF testing and SDF&gt;30% were recruited. SDF test done with SCSA method and randomized using software. ICSI was the method of insemination. Extended embryo culture till blastocyst was done and freeze all policy was opted. Two Blastocysts that showed 100% survival were transferred in a Frozen Embryo transfer (FET) cycle. Embryonic and Reproductive outcomes were compared between both groups. Live birth and Miscarriage were the primary outcomes. Main results and the role of chance Reproductive Outcomes of MACS Vs TESA were: Average Blastocyst conversion - 32% Vs 39% (RR 1.22, CI1.00 to 1.50) Implantation rate (IR) - 50% Vs 35% (RR - 0.71, CI 0.51 to 0.98) Miscarriage rate (MR) - 5.3% Vs 11% (RR1.6333, CI 0.5227 to 5.1039) Multiple Pregnancy rate (MPR) - 8% Vs 4% Live birth Rate (LBR) per Intention to treat (ITT) - 41.3% Vs 44% (RR 0.95, 95% CI 0.72 to 1.26) LBR per ET cycle - 63% Vs 56% (RR 1.23, 95% CI 0.77 to 1.94) Our preliminary results suggest that despite greater availability of blastocysts for transfer in the TESA group, no difference in ART outcomes was observed between the groups. Though the IR was statistically low with TESA, our primary outcomes LBR and MR were comparable. TESA or MACS seem to offer similar outcomes. Considering the invasiveness with TESA, MACS can be offered for better sperm selection for couples with raised sperm DFI & failed implantation. Limitations, reasons for caution Small sample size. TESA is a surgical intervention Wider implications of the findings Optimal intervention for management of SDF still needs further research. Trial registration number CTRI/2019/07/020140

P–084 Microfluidic Sperm Sorting (MFSS) technique versus Physiological Intracytoplasmic Sperm Injection (PICSI) technique in high DNA fragmentation index sperm samples

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Kant ◽  
K D Nayar ◽  
H Sharma ◽  
S Gupta ◽  
S Mishra ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Dna Fragmentation ◽  
Embryo Development ◽  
Intracytoplasmic Sperm Injection ◽  
Development Rate ◽  
Clinical Pregnancy ◽  
Sperm Dna Fragmentation ◽  
Fragmentation Index ◽  
Sperm Dna ◽  
Dna Fragmentation Index

Abstract Study question To evaluate the effectiveness of using Microfluidic Sperm Sorting (MFSS) technique and Physiological Intracytoplasmic Sperm Injection (PICSI) technique in patient with high DNA fragmentation index (DFI) sperm samples. Summary answer Sperm selected by microfluidic sorting are associated with significant increase in day 3 grade A embryo development rate, clinical pregnancy rate over PICSI. What is known already DNA damage is unrecognisable in living sperm prior to insemination and an increased sperm DNA fragmentation index has been associated with lower fertilization rates, impaired embryo development and reduced pregnancy rates. Standard semen processing techniques are associated with centrifugation, which may induce reactive oxygen species and DNA damage. In strategies to minimize sperm DNA fragmentation, Physiological ICSI can relatively reduce sperm DNA fragmentation by 67.9% (Parmegiani et al., 2010) while new technique Microfluidic sperm sorter technique also demonstrate sperm selection with significantly reduced DNA damage. Study design, size, duration A prospective randomised study was conducted from 1st August 2019 to 31st December 2020. Two hundred patients were randomised by computer generated list and divided into 2 groups. Group A (n = 100) , in which sperm were processed by microfluidic sperm sorter (MFSS) while in group B (n = 100), sperm were selected by Physiological Intracytoplasmic Sperm Injection (PICSI) technique and morphologically normal motile sperm were injected by Intracytoplasmic sperm injection (ICSI) technique in all mature oocytes. Participants/materials, setting, methods The study period included all normozoospermic patients with high DNA fragmentation index (&gt;25% ) while oligospermic, asthenozoospermic samples, patients with poor ovarian reserve and advanced age were excluded from the study. All A grade embryos were vitrified and transferred in frozen embryo replacement cycle. Both groups were compared on the basis of fertilisation rate, day 3 grade A embryo development rate , clinical pregnancy rate and miscarriage rate. Main results and the role of chance Cycle characteristics (female age, length of stimulation, gonadotrophin dose, number of oocytes and number of transferred embryos) were similar in both groups. Between the 2 groups, There was a significant increase observed in day 3 grade A embryo development rate (60% vs. 42%, p–0.016) and clinical pregnancy rate (62% vs. 46%, p–0.049), while no statistical significant difference observed in fertilisation rate (82% vs. 78%, p–0.80) and miscarriage rate ( 12% vs. 11%, p- 1). Limitations, reasons for caution: Larger randomised control studies are needed to strengthen these results. Wider implications of the findings: We have demonstrated that sperm sorted by microfluidic helps in selection of sperm with better DNA integrity over Physiological ICSI. Using it in routine practice can help in reducing the negative effect of reactive oxygen species and thus improve pregnancy rate and live birth rate. Trial registration number MCDH/2019/31

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