Colloidal centrifugation with Androcoll-E™ prolongs stallion sperm motility, viability and chromatin integrity

Johannisson, Anders; Morrell, Jane M.; Thorén, Josefina; Jonsson, Magni S.; Dalin, Anne-Marie; Rodriguez‐Martinez, Heriberto

doi:10.1016/j.anireprosci.2009.01.008

Cited by 95 publications

(98 citation statements)

References 30 publications

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“…Aliquots were taken for evaluation of sperm morphology as previously described (Johannisson et al, 2009). Briefly, aliquots (5 μl) were used to prepare air-dried slides for the assessment of sperm head shape of 500 spermatozoa at 1000× magnification.…”

Section: Morphologymentioning

confidence: 99%

“…Reactive oxygen species in stallion semen doses Plasma membrane integrity Assessment of plasma membrane integrity was made using the procedure described by Johannisson et al (2009) at 0 and 2 h after arrival of semen at the laboratory. Briefly, 300 μl of each sample was stained with 0.6 μl of SYBR-14 (final stain concentration 0.4 μM) followed by 3 μl of PI (final stain concentration 24 μM; Live-Dead ® Sperm Viability Lit L-7011; Invitrogen, Eugene, OR, USA).…”

Section: Morphologymentioning

confidence: 99%

“…Sperm chromatin structure assay (SCSA) The method, using acridine orange (AO) and flow cytometric analysis (FCM), was a modification of the method reported in detail previously (Evenson et al, 2002;Johannisson et al, 2009). Briefly, the sperm aliquots were extended with Tris-NaCl-ethylenediaminetetraacetic acid (TNE) buffer (0.15 M NaCl, 0.01 M Tris-HCl, 1 mM EDTA (ethylenediaminetetraacetic acid, pH 7.4) to a final sperm suspension of ∼2 × 10 7 cells/ml.…”

Section: Rosmentioning

confidence: 99%

“…For the purpose of this project, only the %DFI (DNA Fragmentation Index, the percentage of cells outside the main population in the alpha-t-histogram) was included in the statistical analysis. The modification from the method reported previously (Johannisson et al, 2009) was that the frozen-thawed sperm/TNE buffer aliquots were further diluted with TNE buffer (1 : 9 v/v) before adding the detergent solution and the AO.…”

Section: Rosmentioning

confidence: 99%

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Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination

Johannisson

Lundgren

Humblot

et al. 2014

Animal

Self Cite

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The decrease in foaling rates after artificial insemination with cooled semen warrants the search for new predictors of fertility. The objectives were to investigate levels of naturally occurring reactive oxygen species (ROS) in cooled, stored stallion semen doses for artificial insemination (AI), and their relationship with parameters of semen quality and with pregnancy rate. Semen was collected from warmblood stallions (n = 15) and used to prepare commercial semen doses for AI. Sperm quality was evaluated after cooled transport to the laboratory overnight. The results were correlated with observed foaling and pregnancy rates. Hydroethidine and dichlorodihydrofluorescein diacetate were used as indicators for the ROS superoxide and hydrogen pero xide, respectively. Sperm morphology, motility, plasma membrane integrity and chromatin integrity were also evaluated. These variables were correlated with each other and with pregnancy rates. We found a high inter-individual variation in the ROS levels between stallions. The proportion of live, hydrogen peroxide-negative spermatozoa was correlated with progressive motility, whereas live hydrogen peroxide-negative spermatozoa and chromatin damage were negatively correlated, indicating that low levels of hydrogen peroxide were correlated with good chromatin integrity. The percentage of dead hydrogen peroxide-positive sperm was negatively related to the foaling rate. The negative relationships were stronger when combining results from both assays for ROS. These results for stored semen samples indicate that high individual variation exists for superoxide and hydrogen peroxide measurements, and that ROS status can influence sperm quality. Thus, ROS may be some of the factors influencing fertility. Moreover, combinations of ROS variables improved the correlation with fertility, indicating the usefulness of including these variables in a future model for prediction of the fertility of a semen sample.

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Section: Morphologymentioning

confidence: 99%

Section: Morphologymentioning

confidence: 99%

Section: Rosmentioning

confidence: 99%

Section: Rosmentioning

confidence: 99%

See 2 more Smart Citations

Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination

Johannisson

Lundgren

Humblot

et al. 2014

Animal

Self Cite

View full text Add to dashboard Cite

show abstract

“…Single layer colloidal centrifugation of the stallion ejaculate has been shown to result in the selection of the most robust spermatozoa possessing intact and mature chromatin (Johannisson et al, 2009;Morrell et al, 2009aMorrell et al, , 2014bGutiérrez-Cepeda et al, 2011). In addition, potential DNA damage associated with reactive oxygen species (ROS) can be prevented by removing damaged or dead spermatozoa and leukocytes by means of colloidal centrifugation, thus further contributing to maintenance of sperm chromatin integrity (Johannisson et al, 2009;Morrell et al, 2009a,c).…”

mentioning

confidence: 99%

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

Crespo¹,

Gosálvez²,

Johnston³

et al. 2015

Span J Agric Res

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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.Additional key words: Equus caballus; ejaculate fractionation; colloidal centrifugation; sperm DNA fragmentation; DNA longevity; SCD.Abbreviations used: FRAC-1 (first jet, sperm-rich fraction); FRAC-2 (rest of the ejaculate); RAW (combined ejaculate reconstituted post-ejaculation); ROS (reactive oxygen species); rSDF (rate of increase in sperm DNA fragmentation); SCD (sperm chromatin dispersion), SDF (sperm DNA fragmentation).

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A New Method for Evaluating Stallion Sperm Viability and Mitochondrial Membrane Potential in Fixed Semen Samples

Peña

Ball

Squires

2017

Cytometry Part B Clinical

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Multiparametric assessment of stallion sperm quality using flow cytometry can be a useful adjunct in semen evaluation; however, the availability of flow cytometers in veterinary practice is limited. The ability to preserve and transport sperm samples for later flow cytometric analysis using fixable probes would potentially facilitate this process. In the current study, we validated the combination of live/dead Zombie Green (a fixable dye used to assess live and dead sperm) and MitoTracker Deep Red (used to assess mitochondrial membrane potential). The assay was validated against classic, non-fixable, membrane assays (SYBR-14/PI). Our results demonstrated the feasibility of the assay. In conclusion, stained and fixed semen samples stored for 72 h obtained equivalent results to the exam on the same day; this new protocol shall facilitate the wider use of flow cytometry in stallion andrology in the future. © 2017 International Clinical Cytometry Society.

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Colloidal centrifugation with Androcoll-E™ prolongs stallion sperm motility, viability and chromatin integrity

Cited by 95 publications

References 30 publications

Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination

Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination

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

A New Method for Evaluating Stallion Sperm Viability and Mitochondrial Membrane Potential in Fixed Semen Samples

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