Comparison of different methods of goat sperm selection and capacitation for optimization of assisted reproductive technologies

Olivares, C. C. S.; Fonseca, J. F. da; Camargo, L. S. A.; Souza-Fabjan, J. M. G.; Rodrigues, A. L. R.; Brandão, Felipe Zandonadi

doi:10.1016/j.smallrumres.2015.04.009

Cited by 10 publications

(9 citation statements)

References 27 publications

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“…Lower recovery rate was obtained after swimup, which is relatively low and corroborate with several other studies comparing different sperm selection techniques on bovine, ovine and caprine [24,27,30,36,37]. In this study, Percoll and mini-Percoll recovered approximately 10 times more cells than swim-up, which is an important feature to be considered during sperm preparation for IVF, being possible to use only one semen straw.…”

Section: Discussionsupporting

confidence: 89%

Comparison of Different Sperm Selection Techniques in Ram Frozen-Thawed Sperm

Olivares

Souza-Fabjan

Fonseca

et al. 2017

Acta Scientiae. Vet.

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Background: The success of fertilization is directly associated with semen quality and the sperm preparation. Considering the common use of cryopreserved spermatozoa, there is a need to develop strategies for sperm preparation in order to achieve a sperm sample of high quality through a rigorous selection of sperm. Thus, sperm cells are being more extensively investigate. This study aimed evaluating the influence of different sperm selection techniques on ram sperm parameters in semen preparation.Materials, Methods & Results: Frozen-thawed commercial semen from 10 Santa Inês rams was subjected to either: swim-up, Percoll, mini-Percoll, sperm washing by centrifugation or a control group. After each technique, samples were incubated at 37°C for 1 h, 2 h and 3 h. At post-selection moment (0 h) and at each interval, sperm recovery rate, motility, capacitation and plasma membrane (PM) integrity were analyzed. The lowest (P < 0.05) recovery rate was recorded after swim-up (1.0 ± 0.3%), whilst the others were similar (P > 0.05). Most part of motility parameters were not affected (P > 0.05) by the technique at 0 h; just swim-up obtained higher (P < 0.05) values for VSL (41.8 ± 11.1) and VAP (46.9 ± 11.2). Overall, swim-up presented higher (P < 0.05) values for most of motility parameters over time of incubation. The control group led to more (P < 0.05) capacitated cells (46.8 ± 2.3%), whilst Percoll, mini-Percoll and swim-up were similar (~33%; P > 0.05), regardless of interval incubation. However, the latter three techniques presented more (P < 0.05) reacted cells. Swim-up recovered a higher (P < 0.05) number of intact cells (32.1 ± 6.4%), and Percoll, mini-Percoll and control group were similar (P > 0.05).Discussion: The present study evaluated the sperm cell and its preparation for receiving the oocyte under optimal conditions. evaluate the sperm cell and its preparation for receiving the oocyte under optimal conditions. Although the swim-up technique promoted higher rates for some of the sperm parameters evaluated, Percoll protocols are the most widely used procedures for selection during the sperm preparation in many species, possibly because of its greater speed, practical method and convenience compared to the swim-up technique. Percoll and mini-Percoll recovered approximately 10 times more cells than swim-up, which is an important feature to be considered during sperm preparation for in vitro fertilization (IVF), being possible to use only one semen straw. The high capacitated and acrosome reaction rates observed after the treatments in the current study, are probably reinforced by changes in sperm cells caused by the cryopreservation process. In order to strength the evidence that frozen-thawed sperm, even after selection, is sensible and reactive to capacitation-like events, we demonstrated the capacitated and acrosome-reacted cell values immediately after the selection treatments behaved differently than when authors used ram fresh sperm. Possibly, this capacitation-like changes observed in frozen-thawed sperm occurs regardless of the selection treatment used. Analyzing the motility parameters immediately after the selection, all treatments maintained or increased the rates compared to the control group. The swim-up, mini-Percoll and Percoll did not differ in any parameters. Given that Percoll and mini-Percoll did not show differences in relation to swim-up for motility parameters, such techniques can be used to replace the latter, obtaining similar sperm samples with good quality. However, swim-up technique involves a procedure that recovers a clean fraction without debris and other types of cells, with high rate of mobile sperm with excellent quality, reason why it can justify the higher recovery of intact spermatozoa after the technique.

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Section: Discussionsupporting

confidence: 89%

Comparison of Different Sperm Selection Techniques in Ram Frozen-Thawed Sperm

Olivares

Souza-Fabjan

Fonseca

et al. 2017

Acta Scientiae. Vet.

Self Cite

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“…Parameters such as sperm volume, sperm motility, morphology, and concentration are commonly used to achieve this qualification. The production of ROS by spermatozoa is a normal physiological process necessary for the occurrence of sperm capacitation and the acrosomal reaction (Olivares et al, 2015). However, long periods of storage in the epididymis can generate oxidative stress and the consequent DNA damage, as well as lipid peroxidation of the sperm membrane, leading to a reduction in sperm motility (Silva, 2011).…”

Section: Discussionmentioning

confidence: 99%

Seminal quality comparison of first and second cryopreserved ejaculates of Alpine kid goats by flow cytometry

Novaes

Almeida

Netto

et al. 2022

Zygote

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Summary Discarding the first ejaculate is recommended as an alternative for improving seminal quality after long sexual resting, especially when semen should be used for cryopreservation. However, when the males are not in sexual resting the necessity to discarding the first ejaculate is still unknown. Therefore, this study aimed to compare by flow cytometry the quality of the first and second ejaculates. Ten kids and uniform goats between 5 and 6 months of age were used in a completely randomized design. Semen collection was carried out every 4 days, until a total of five ejaculates per animal in each treatment was completed. The fresh and frozen semen collected were processed and analyzed using macroscopic and microscopic parameters, resistance test, hypo-osmotic medium test, and flow cytometry (FC). The FC parameters were production of reactive oxygen species, plasma and acrosomal membrane integrity, and lipid peroxidation of the plasma membrane. The ejaculates did not differ for the resistance test, the reactivity in the hypo-osmotic medium and for the macroscopic and microscopic seminal parameters, except for sperm volume and concentration. The first ejaculate had a higher percentage of minor and total defects. None of the FC parameters analyzed differed between the first and second ejaculates. The first and second ejaculates demonstrated similar seminal qualities, so for Alpine kid goats without a sexual resting period, discarding the first ejaculate it is not recommended.

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“…The sperm was selected by the mini-Percoll technique in accordance with the method we previously established in sheep [15]. The gradient was formed by pipetting 0.4 mL of 90% Percoll solution into a 1.5 mL microtube and then overlaying it with 0.4 mL of 45% Percoll solution.…”

Section: Mini-percoll Techniquementioning

confidence: 99%

Mini-Percoll Technique Induces Similar Capacitation Features in Domestic Ruminant Frozen-Thawed Spermatozoa Regardless of the Presence of Heparin

Alfradique

Olivares

Souza-Fabjan

et al. 2019

Acta Scientiae. Vet.

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Background: Sperm capacitation is a process consists of a series of functional, biochemical, and biophysical modifications that render the ejaculated sperm competent for oocyte fertilization. Secreted by the female reproductive tract epithelium, heparin promotes capacitation by binding to and removing seminal plasma proteins, which are adsorbed to the sperm PM and would inhibit capacitation. There is substantial evidence that cryopreservation promotes capacitation-like changes in bull, ram and buck sperm. Our general hypotheses were: (a) cryopreserved ram sperm suffer capacitation more quickly than buck and bull sperm under the same conditions; (b) the capacitation status of ruminant cryopreserved sperm is similar whether or not heparin is present after the mini-Percoll technique; and (c) ruminant frozen-thawed sperm selected by mini-Percoll and incubated within media without heparin supplementation is not impaired in terms of capacitation status and sperm agglutination. This study aimed to compare sperm parameters of ovine, caprine, and bovine frozen-thawed sperm after mini-Percoll processing followed by incubation with or without heparin supplementation.Materials, Methods & Results: Commercial semen of all species were used. Sperm samples were selected by mini-Percoll and supplemented (or not) with heparin within an incubation medium for 18 h. Sperm kinematics (CASA system analyzes), capacitation status (CTC staining) and sperm agglutination were evaluated after thawing, mini-Percoll, 1.5 h, 3 h, 6 h and 18 h. In comparison with post-thawing analysis, ovine species demonstrated a reduction (P < 0.05) in most of the sperm motility parameters after mini-Percoll. Conversely, ovine samples presented the highest (P < 0.05) rate of acrosome-reacted cells after mini-Percoll. Heparin supplementation did not affect most of the parameters evaluated (P > 0.05). In caprine and bovine species, a lower (P < 0.05) rate of sperm agglutination was observed in the presence of heparin at 18 h of incubation. In the absence of heparin, ovine samples showed a higher (P < 0.05) agglutination rate compared to the bovine species after long incubation period.Discussion: The present study compared sperm parameters (sperm kinematics, agglutination rate and capacitation status) of ruminant frozen-thawed sperm after mini-Percoll selection followed by in vitro incubation with or without heparin supplementation. In this study, it was observed the same rate of capacitated cells after the sperm selection (min-Percoll) between ruminant species. This indicate that the capacitation process occurs similarly between ruminant species, refuting the first hypothesis of this study. The presence of heparin did not influence the capacitation status of ruminant frozen-thawed sperm after mini-Percoll selection, it demonstrates that the second hypothesis was supported by this study making more economic and practical the use of ruminant frozen-thawed semen. The absence of heparin in the incubation medium did not harmed the capacitation status and sperm agglutination of ruminant frozen-thawed sperm. This supported the third hypothesis of the current study and indicate that the use of mini-Percoll technique regardless the presence of heparin could be a useful alternative for the preparation of ruminant frozen-thawed sperm. In conclusion, the capacitation status of ruminant frozen-thawed sperm is similar whether or not heparin is present after the mini-Percoll technique.

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Comparison of different methods of goat sperm selection and capacitation for optimization of assisted reproductive technologies

Cited by 10 publications

References 27 publications

Comparison of Different Sperm Selection Techniques in Ram Frozen-Thawed Sperm

Comparison of Different Sperm Selection Techniques in Ram Frozen-Thawed Sperm

Seminal quality comparison of first and second cryopreserved ejaculates of Alpine kid goats by flow cytometry

Mini-Percoll Technique Induces Similar Capacitation Features in Domestic Ruminant Frozen-Thawed Spermatozoa Regardless of the Presence of Heparin

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