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>

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.

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

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.

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