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Reproductive performance of sows inseminated with semen doses stored for up to seven days in long-term extender in a field condition.

ABSTRACT: This study aimed to evaluate the reproductive performance of sows inseminated with semen doses preserved at 15-18 °C for up to seven days in long-term extender (Duragen®). Parity one (PO1) to PO7 sows were randomly assigned to the following groups: AI1-3 (n=190), insemination with semen doses stored between one and three days; and AI5-7 (n=124), insemination with semen doses stored between five and seven days. Sows were submitted to estrus detection twice a day. Post-cervical insemination according to weaning-to-estrus interval was performed. The farrowing rate (FR) did not differ between the groups (AI1-3=83.2%; AI5-7=82.2%; p>0.05) nor did the total number of piglets born (TPB; AI1-3=14.2±0.3; AI5-7=14.5±0.3; p>0.05). Considering the semen dose most likely responsible for fertilization according to its storage time (1, 2-3, 5, and 6-7 days), the FR (72.7%, 87.8%, 85.7%, and 79%, respectively) and TPB (14.4, 14.0, 14.9, and 13.5, respectively) were similar among the groups (p>0.05). In conclusion, the use of semen doses extended with long-term extender stored for up to seven days did not impair the reproductive performance of sows. Therefore, it's using could optimize production efficiency and logistics of semen dose deliveries to sow farms.

SUBMITTER: Lucca MS

PROVIDER: S-EPMC7189442 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Reproductive performance of sows inseminated with semen doses stored for up to seven days in long-term extender in a field condition.

Lucca Matheus Schardong MS Ulguim Rafael da Rosa RDR Bortolozzo Fernando Pandolfo FP Wentz Ivo I Rocha Janaina Colecha JC de Souza Mayara Corrêa Dias MCD Escobar Roberto Vasconcelos RV Calderam Kérlin K de Quadros Pedro Ivo PI Marcos Raquel Ausejo RA

Animal reproduction 20200211 1

This study aimed to evaluate the reproductive performance of sows inseminated with semen doses preserved at 15-18 °C for up to seven days in long-term extender (Duragen<sup>®</sup>). Parity one (PO1) to PO7 sows were randomly assigned to the following groups: AI1-3 (n=190), insemination with semen doses stored between one and three days; and AI5-7 (n=124), insemination with semen doses stored between five and seven days. Sows were submitted to estrus detection twice a day. Post-cervical insemina ...[more]

PMID: 32368281

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Project description:IntroductionSemen cryopreservation is the most popular practice for semen production for artificial insemination and in vitro fertilization in cattle. The Seminal plasma contains extracellular vesicles (spEVs) which modulate sperm viability and function during oocyte fecundation. The study of spEVs in frozen-thawed semen doses may yield novel indicators for predicting bull fertility, but the presence of the semen extender may hinder molecular profiling of spEVs. The aim of this study was to provide extensive characterization of EVs isolated from seminal plasma before and after the cryopreservation process and the addition of a commercial animal protein-free semen extender to understand the potential influence of EVs originating from the extender in hindering the use of spEVs derived biomarkers for assessment of bull fertility.MethodsEVs were isolated from the seminal plasma (with or without the extender), from the cryopreserved straw devoid of spermatozoa, and from the extender using two different methods, ultracentrifugation (UC) and size exclusion chromatography (SEC), and characterized for their structure and composition.ResultsPhysical characterization of EVs showed that size and particle numbers were related to the method of isolation. spEVs were larger but less abundant (UC: 168.9 nm, n = 2.68 × 109; SEC: 197.0 nm, n = 6.42 × 109) compared to extender EVs (UC: 129.0 nm, n = 2.68 × 1011; SEC: 161.8 nm, n = 6.47 × 1011). Western blotting analysis (WB) confirmed the presence of typical EV markers in spEVS: the membrane bound CD9 (25 kDa) and the luminal markers Alix (96 kDa) and TSG101 (48 KDa). Although Transmission Electron Microscopy confirmed the presence of a lipid bilayer structure in all preparations, no specific EV markers were detected in the vesicles isolated from extender when the Single Molecule Array (SiMoa) was used. A total of 724 Bos taurus miRNAs were identified in at least one preparation. The percentage of miRNAs identified in EVs from the extender (0.05%-0.49% of the total reads) was lower than in the preparation containing spEVs (10.56%-63.69% of the total reads). Edge-R identified a total of 111 DE-miRNAs between EVs isolated from the extender by two methods. Among them, 11 DE-miRNAs (bta-miR-11980, bta-miR-11987, bta-miR-12057, bta-miR-1246, bta-miR-125b, bta-miR-181b, bta-miR-2340, bta-miR-2358, bta-miR-2478, bta-miR-2898, and bta-miR-345-3p) were also abundant in EVs isolated from seminal plasma preparations with extender.ConclusionThis study clearly demonstrates that the presence of the extender does not prevent the characterization of spEVs in cryopreserved semen. However, the molecular profiling of spEVs can be influenced by the isolation method used and by the presence of some miRNAs from the extender. Therefore, in such studies, it is advisable to characterize both spEVs and the vesicles isolated from the extender.

Project description:The long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation's success rate. The aim of our study was to reduce the percentage of glycerol in the extender by elimination or substitution with biocompatible, non-toxic polysaccharides. For boar semen extender improvement, we tested a novel modification with the polysaccharides dextran and pentaisomaltose in combination with unique in silico predictive modeling. We targeted the analysis of in vitro qualitative sperm parameters such as motility, viability, mitochondrial activity, acrosome integrity, and DNA integrity. Non-penetrating polysaccharide-based cryoprotective agents interact with sperm surface proteins such as spermadhesins, which are recognized as fertility markers of boar sperm quality. The in silico docking study showed a moderate binding affinity of dextran and pentaisomaltose toward one specific spermadhesin known as AWN, which is located in the sperm plasma membrane. Pentaisomaltose formed a hydrophobic pocket for the AWN protein, and the higher energy of this protein-ligand complex compared with dextran was calculated. In addition, the root mean square deviation (RMSD) analysis for the molecular dynamics (MD) of both polysaccharides and AWN simulation suggests their interaction was highly stable. The in silico results were supported by in vitro experiments. In the experimental groups where glycerol was partially or entirely substituted, the use of pentaisomaltose resulted in improved sperm mitochondrial activity and DNA integrity after thawing when compared with dextran. In this paper, we demonstrate that pentaisomaltose, previously used for cryopreservation in hematopoietic stem cells, represents a promising compound for the elimination or reduction of glycerol in extenders for boar semen cryopreservation. This novel approach, using in silico computer prediction and in vitro testing, represents a promising technique to help identify new cryoprotectants for use in animal breeding or genetic resource programs.

Dataset Information

Reproductive performance of sows inseminated with semen doses stored for up to seven days in long-term extender in a field condition.

Publications

Reproductive performance of sows inseminated with semen doses stored for up to seven days in long-term extender in a field condition.

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