A new test based on the hypotonic resistance and functional competence to evaluate the sperm quality, cryotolerance and in vitro fertilizing ability in pigs

2019 ◽  
Vol 140 ◽  
pp. 84-92
Author(s):  
Julian Valencia ◽  
Marc Yeste ◽  
Armando Quintero-Moreno ◽  
Francisco Javier Henao
Keyword(s):  
Sperm Quality ◽  
Functional Competence ◽  
Fertilizing Ability

scholarly journals Validation of the Turkey Semen Cryopreservation by Evaluating the Effect of Two Diluents and the Inseminating Doses

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1329
Author(s):  
Michele Di Iorio ◽  
Giusy Rusco ◽  
Roberta Iampietro ◽  
Lucia Maiuro ◽  
Achille Schiavone ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
In Vivo Test ◽  
Sperm Analysis ◽  
Fertilizing Ability ◽  
Vitro Analysis ◽  
Hatching Rates

This study was designed to test the fertilizing ability of cryopreserved turkey semen, and here, two experiments were performed: an in vitro analysis to assess the effects of Tselutin and Lake diluents and an in vivo test to determine the fertility and hatching rates by also studying the feat of three insemination doses (250, 400 and 600 × 106 sperm/hen). Pooled semen samples were diluted with Tselutin or Lake extender which contained 20% of dimethylsulfoxide and 1 mM of Ficoll at final sperm concentration of 3 × 109 sperm/mL. Thereafter, semen was packaged into straws and frozen on liquid nitrogen. The post-thaw sperm quality was evaluated considering motility (computer-aided sperm analysis—CASA system) and membrane integrity (flow cytometry). Significantly higher values of progressive motility and some kinetic parameters in semen frozen with Lake were found. When we compared the extenders in vivo, no significant effects were detected, whilst sperm concentration significantly affected both fertility and hatching rates, with the best results obtained with the sperm concentration of 400 × 106 sperm/hen. From the results obtained, it emerged that the extender type only affected sperm motility characteristics, not the fertilizing ability of frozen-thawed semen, while inseminating dose markedly affected fertility and hatching rates.

151 Effect of season on the in vitro fertilizing ability of frozen - thawed bovine spermatozoa

2019 ◽  
Vol 31 (1) ◽  
pp. 200
Author(s):  
M. Sabes-Alsina ◽  
M. Wallgren ◽  
Y. Sjunesson ◽  
N. Lundeheim ◽  
M. López-Béjar ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Mitochondrial Potential ◽  
Semen Collection ◽  
Frozen Semen ◽  
Fertilizing Ability

Previous research indicated that the season during which oocytes are harvested affects quality of in vitro-produced embryos (Gupta et al. 2016 Anim. Reprod. Sci. 164, 162). In our own studies, sperm kinematics, membrane integrity, acrosome status, mitochondrial potential and reactive oxygen species were affected by season of semen collection (Sabés-Alsina et al. 2017 Vet. Rec. 180, 251). The aim of this study was to investigate effect of season of semen collection on in vitro fertilizing ability and embryo development of the same sperm samples from the sperm quality study, using lower than normal sperm doses to detect small differences between groups (Ward et al. 2003 Theriogenology 59, 1575). Frozen semen was available from 8 Holstein bulls kept outdoors in northern Spain, collected during winter, spring and summer. Bovine ovaries, Holstein and Swedish Red breeds, were obtained from an abattoir in spring. Oocytes were matured and fertilized in vitro with a low dose of frozen-thawed sperm (sperm:oocyte ratio 2500:1). After fertilization, presumptive embryos were evaluated at 44h for cleavage and on Day 8 for blastocyst development. Number of sperm binding to the zona pellucida and number of nuclei in developing blastocysts were assessed after staining with Hoechst 33342. There were 2 or 3 replicates per bull. For 555 oocytes inseminated, cleavage rates for winter, spring and summer semen collections were 42, 49 and 47%, respectively; blastocyst rates were 7, 12 and 8%; blastocyst cell numbers were 8.7, 10.2 and 7.8; and mean number of sperm bound was 0.70, 0.63 and 0.50. Although there were no differences (P>0.05) due to season of semen collection for any of these end points, individual bulls had considerable variation in cleavage rate. In conclusion, despite previously published differences in sperm quality with season from these bulls, ability of frozen-thawed sperm to fertilize bovine oocytes and initial embryo development in vitro were not affected by season of semen collection, at least for oocytes collected in spring. However, bull-to-bull variation in cleavage rate was high. The authors gratefully acknowledge the financial support of the Ministerio de Economía y Competitividad and FEDER (AGL2016-79802-P). M. Sabes-Alsina was supported by a PIF from the Universitat Autònoma de Barcelona and by the STS of the Epiconcept COST Action; J. M. Morrell was funded by the Swedish Research Council for the environment, agricultural sciences and spatial planning (FORMAS; 221-2010-1241) and the Swedish Farmers’ Association (SLF; 1330039).

scholarly journals Improving the Rabbit Semen Cryopreservation Protocol: Comparison Between Two Extenders and Inseminating Doses

2020 ◽  
Vol 20 (3) ◽  
pp. 887-898
Author(s):  
Michele Di Iorio ◽  
Giusy Rusco ◽  
Maria Antonietta Colonna ◽  
Michele Schiavitto ◽  
Maria Silvia D’Andrea ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Artificial Insemination ◽  
Sperm Quality ◽  
Dna Integrity ◽  
Semen Cryopreservation ◽  
Fertilizing Ability ◽  
Cryopreservation Protocol

AbstractThis study has been designed to optimize the semen freezing protocol in rabbits, in this regard we compared a Tris-citrate-glucose (TCG) extender with a commercial one (Cortalap®), that to the best of our knowledge has never been used up to now on the in vitro freezability and fertilizing ability of cryopreserved rabbit semen. Two different inseminating semen doses were considered. Five pooled semen samples were divided into two subsamples and each of them were diluted to a ratio 1:1 (v:v) with a freezing extender composed of TCG or Cortalap® containing 16% of dimethylsulfoxide and 0.1 mol/L of sucrose. The extended semen was filled into 0.25 mL plastic straws and frozen above a liquid nitrogen surface. After thawing (50°C/10 seconds) we determined sperm motility, viability, membrane functionality, acrosome and DNA integrity. Our results showed that the Cortalap® extender significantly improved the in vitro post-thaw sperm quality, in comparison to TCG. When we compared the extenders in vivo, no significant differences in the reproductive performances were observed independently by inseminating doses used. In this study we demonstrated that Cortalap® extender can be used as an alternative to TCG. Thus, the Cortalap® being a ready to use extender, implies a reduction of time, mistakes and microbial contaminations during its preparation. This discovery results as significant because it provides beyond an important contribution to the creation of the first Italian semen cryobank of rabbit breeds and also for livestock rabbit farms based on artificial insemination (AI) program.

Influence of in vitro exposure to mycotoxin zearalenone and its derivatives on swine sperm quality

2008 ◽  
Vol 25 (4) ◽  
pp. 461-467 ◽  
Author(s):  
E. Benzoni ◽  
F. Minervini ◽  
A. Giannoccaro ◽  
F. Fornelli ◽  
D. Vigo ◽  
...  
Keyword(s):  
Sperm Quality ◽  
In Vitro Exposure

Seminal plasma proteins protect flow-sorted ram spermatozoa from freeze - thaw damage

2009 ◽  
Vol 21 (4) ◽  
pp. 571 ◽  
Author(s):  
T. Leahy ◽  
J. I. Marti ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Seminal Plasma ◽  
High Speed ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Weight Fraction ◽  
Protein Supplementation ◽  
Freezing Process ◽  
Spermatozoa Motility ◽  
Functional Integrity

Seminal plasma improves the functional integrity of compromised ram spermatozoa but has been reported to be toxic to sorted spermatozoa. The present study attempted to clarify this paradoxical effect and improve the functional integrity of spermatozoa following sorting and cryopreservation. The in vitro function of sorted spermatozoa (motility characteristics and membrane integrity) was examined after supplementation with differing concentrations and protein fractions of seminal plasma at various stages of the sorting and freezing process. For all experiments, spermatozoa (two males, n = four ejaculates per male) were processed through a high-speed flow cytometer before cryopreservation, thawing and incubation for 6 h (37°C). Supplementation of crude seminal plasma (CP), its low molecular weight fraction (LP; <10 kDa) or protein-rich fraction (SPP; >10 kDa), immediately before freezing improved the functional integrity of sorted spermatozoa compared with no supplementation (control), whereas supplementation after thawing had no effect for CP and LP. The protective effect of seminal plasma was not altered by increasing the amount of protein supplementation. No toxic effect of CP, SPP or LP was evident even when supplemented at high protein concentrations. It is concluded that seminal plasma protein, if added to ram spermatozoa after sorting and before freezing, can improve post-thaw sperm quality and consequently the efficiency of sorting. This effect is most likely related to protection of the spermatozoa during freeze–thawing.

scholarly journals Post-wash total motile sperm count a useful predictor in the decision to perform IVF/ICSI in patients with non-male factor infertility

2020 ◽  
Vol 3 (2) ◽  
pp. 99-106
Author(s):  
Sara Mahmood Qureshi ◽  
Salma Kafeel ◽  
Riffat Bibi ◽  
Jawad Mohmand
Keyword(s):  
Sperm Quality ◽  
Sperm Count ◽  
Adverse Outcomes ◽  
Male Factor Infertility ◽  
Motile Sperm ◽  
Operating Characteristics ◽  
Male Factor ◽  
Vitro Fertilization ◽  
Total Motile Sperm Count

Introduction: The unrestricted use of intracytoplasmic sperm injection (ICSI) for non-male factor infertility is associated with adverse outcomes. Post-wash total motile sperm count (PW-TMSC) offers prognostic value to assess sperm quality and aid in the decision to perform in vitro fertilization (IVF) or ICSI. Objectives: The aim of this study was to identify the effect of PW-TMSC on fertilization rates in patients undergoing IVF cycles exclusively with non-male factor infertility. It also aimed to identify whether unnecessary ICSI could be avoided in such cases, thus maximizing optimal outcomes. Materials & Methods: We retrospectively analyzed age, semen volume, prewash TMSC, and PW-TMSC in 68 conventional IVF cycles of infertile couples with non-male factor infertility. Clinical characteristics including female age, number of follicles, level of estradiol on trigger day, mature cumulus-oocyte complexes (COCs) collected, were also included. Results: Incidence of <30% fertilization was significantly higher in the 4-<10 Million group compared with the ≥20 Million post-wash TMSC group (P<0.001). Furthermore, Receiver operating characteristics (ROC) analysis revealed post-wash TMSC as a significant predictor (P<0.05) of total failed fertilization (TFF) and of ≥30% fertilization (P<0.05) with area under curve (AUC) of 0. 79 and 0.77, respectively, with a deemed cutoff of 10.89 Million. Conclusion: Post-wash TMSC is a good predictor of fertilization; it can help in avoiding potentially low or even total fertilization failure (TFF). A cut-off point of 10.89 Million or less should warrant the use of ICSI.

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