scholarly journals Tolerance of Stored Boar Spermatozoa to Autologous Seminal Plasma: A Proteomic and Lipidomic Approach

2020 ◽  
Vol 21 (18) ◽  
pp. 6474
Author(s):  
Lisa Höfner ◽  
Anne-Marie Luther ◽  
Alessandra Palladini ◽  
Thomas Fröhlich ◽  
Dagmar Waberski
Keyword(s):  
Mass Spectrometry ◽  
Seminal Plasma ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Protein Markers ◽  
Long Term Storage ◽  
Lipidome Analysis ◽  
Boar Spermatozoa ◽  
Term Storage

Long-term exposure of liquid preserved boar spermatozoa to seminal plasma (SP) can cause dramatic sperm injury. This study examined whether boar specificity exists in the sensitivity of spermatozoa to SP and whether correspondent biomarkers can be identified. Consecutive ejaculates (n = 4–5) collected from 19 boars were centrifuged, diluted with a pH-stablising extender with 10% (v/v) autologous SP and evaluated by computer-assisted semen analysis and flow cytometry. Up until 144 h storage, four boars showed consistently high sperm motility, viability and mitochondria activity, and one boar showed consistently low values. Intra-boar variability was high in the other boars. Screening of SP (n = 12 samples) for protein markers using mass spectrometry identified three protein candidates of which the granulin precursor, legumain and AWN were 0.5 to 0.9 log2-fold less abundant (p < 0.05) in SP-resistant compared to SP-sensitive samples. Lipidome analysis by mass spectrometry revealed 568 lipids showing no difference between the SP-groups. The most abundant lipids were cholesterol (42,442 pmol), followed by phosphatidylserine (20,956 pmol) and ether-linked phosphatidylethanolamine (13,039 pmol). In conclusion, three candidate proteins were identified which might be indicative of SP-tolerance of sperm during long-term storage. Noteworthy, a first lipidomic profile of boar SP is presented.

201 EFFECTS OF REMOVING SEMINAL PLASMA DURING 8-h STORAGE BEFORE SEX-SORTING BOVINE SPERM

2012 ◽  
Vol 24 (1) ◽  
pp. 213 ◽  
Author(s):  
C. A. Burroughs ◽  
K. M. Evans ◽  
R. W. Lenz ◽  
G. E. Seidel
Keyword(s):  
Seminal Plasma ◽  
Sperm Concentration ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Long Term Storage ◽  
Bovine Sperm ◽  
Standard Procedures ◽  
Term Storage ◽  
Artificial Vagina

We evaluated sex-sorting parameters and post-thaw motility for sperm stored with or without seminal plasma for 8 h before sorting. One first ejaculate was collected from each of 6 bulls routinely collected via artificial vagina; ejaculates contained at least 70% motile and 75% morphologically normal sperm and sperm concentrations ranged from 0.75 to 2.21 × 109 sperm mL–1. Ejaculates were divided into 2 samples and centrifuged at 1000 × g for 15 min. Seminal plasma from 1 sample was replaced with TALP (pH 7.4) to a sperm concentration of 1.4 × 109 sperm mL–1. The seminal plasma/sperm admixture of the other sample (control) was suspended to initial ejaculate sperm concentration. Both samples were stored for 8 h at 16°C before being subjected to standard sex-sorting procedures. Sperm were analyzed and bulk sorted on a MoFlo SX (XY Inc., Navasota, TX, USA) flow cytometer/cell sorter for percentage of live-oriented cells, percentage of membrane-impaired sperm (cell membranes permeable to red food colouring, which were discarded during sorting) and resolution between X- and Y-bearing sperm populations (peak to valley ratio). Sorted sperm were frozen according to standard procedures and post-thaw motility was determined immediately after thawing using computer-assisted sperm analysis. Treatments were compared using a paired t-test. Control sperm stored with seminal plasma resulted in a higher percentage of live-oriented cells (55%) versus those stored without seminal plasma (51%; P = 0.02). The percentage of membrane-impaired sperm was lower for control sperm (19%) than that of samples without seminal plasma (28%; P < 0.001). Resolution was greater for sperm stored without seminal plasma (34%) than for control sperm (10%; P = 0.04). Post-thaw, both total and progressively motile sperm were higher for samples without seminal plasma (63 and 53%, respectively) compared with those of the control samples (52 and 45%, respectively; P < 0.04). In conclusion, sperm stored for 8 h without seminal plasma had greater resolution between X- and Y-bearing populations and higher post-thaw motility than control sperm. However, these samples had a higher percentage of membrane-impaired sperm that were removed during sorting. Long-term storage of sperm in their seminal plasma before sex-sorting appears to be detrimental to post-sorting, post-thaw sperm motility.

scholarly journals Hyaluronan improves neither the long-term storage nor the cryosurvival of liquid-stored CD44-bearing AI boar spermatozoa

2018 ◽  
Vol 64 (4) ◽  
pp. 351-360 ◽  
Author(s):  
Manuel ÁLVAREZ-RODRIGUEZ ◽  
Alejandro VICENTE-CARRILLO ◽  
Heriberto RODRIGUEZ-MARTINEZ
Keyword(s):  
Long Term Storage ◽  
Boar Spermatozoa ◽  
Term Storage

scholarly journals Cryopreservation of boar semen

2020 ◽  
Vol 65 (No. 4) ◽  
pp. 115-123
Author(s):  
Marija Jovičić ◽  
Eva Chmelíková ◽  
Markéta Sedmíková
Keyword(s):  
Animal Species ◽  
Semen Quality ◽  
Egg Yolk ◽  
High Rate ◽  
Freezing And Thawing ◽  
Long Term Storage ◽  
Boar Semen ◽  
Boar Spermatozoa ◽  
Term Storage

Sperm cryopreservation is the best technology for long-term storage of the semen. However, the damage of boar spermatozoa by cryopreservation is more severe than in other animal species and a standardized freezing protocol for efficient cryopreservation has not been established yet. Semen quality and freezability vary greatly between breeds as well as between individual boars and even the season. Boar spermatozoa are sensitive to low temperatures; they sustain damage and a high rate of mortality and freezing/thawing the boar semen may strongly impair the sperm function and decrease the semen quality. The freezability of boar semen can be influenced by a cryopreservation procedure, and also by using various additives to freezing and thawing extenders such as antioxidants. In order to obtain acceptable results after thawing the boar semen, it is necessary to combine an optimal amount of additives (glycerol, egg yolk, sugars, antioxidants), cooling and warming velocities.

scholarly journals Impact of Freeze-thaw Cycles and Storage Time on Plasma Samples Used in Mass Spectrometry Based Biomarker Discovery Projects

2005 ◽  
Vol 1 ◽  
pp. 117693510500100 ◽  
Author(s):  
Breeana L Mitchell ◽  
Yutaka Yasui ◽  
Christopher I Li ◽  
Annette L. Fitzpatrick ◽  
Paul D Lampe
Keyword(s):  
Mass Spectrometry ◽  
Biomarker Discovery ◽  
Plasma Proteome ◽  
Sample Handling ◽  
Freeze Thaw ◽  
Long Term Storage ◽  
And Storage ◽  
Human Fluids ◽  
Term Storage

Mass spectrometry approaches to biomarker discovery in human fluids have received a great deal of attention in recent years. While mass spectrometry instrumentation and analysis approaches have been widely investigated, little attention has been paid to how sample handling can impact the plasma proteome and therefore influence biomarker discovery. We have investigated the effects of two main aspects of sample handling on MALDI-TOF data: repeated freeze-thaw cycles and the effects of long-term storage of plasma at –70°C. Repeated freeze-thaw cycles resulted in a trend towards increasing changes in peak intensity, particularly after two thaws. However, a 4-year difference in long-term storage appears to have minimal effect on protein in plasma as no differences in peak number, mass distribution, or coefficient of variation were found between samples. Therefore, limiting freeze/thaw cycles seems more important to maintaining the integrity of the plasma proteome than degradation caused by long-term storage at –70°C.

Long-Term Storage Facility for Reactor Compartments in Sayda Bay: German Support for Utilization of Nuclear Submarines in Russia

2007 ◽  
Author(s):  
Dietmar Wolff ◽  
Holger Vo¨lzke ◽  
Wolfgang Weber ◽  
Volker Noack ◽  
Gu¨nther Ba¨uerle
Keyword(s):  
Computer Assisted ◽  
Storage Facility ◽  
Nuclear Submarine ◽  
Long Term Storage ◽  
Global Partnership ◽  
Federal Institute ◽  
Almost All ◽  
Interim Storage ◽  
Term Storage

The German-Russian project that is part of the G8 initiative on Global Partnership Against the Spread of Weapons and Materials of Mass Destruction focuses on the speedy construction of a land-based interim storage facility for nuclear submarine reactor compartments at Sayda Bay near Murmansk. This project includes the required infrastructure facilities for long-term storage of about 150 reactor compartments for a period of about 70 years. The interim storage facility is a precondition for effective activities of decommissioning and dismantlement of almost all nuclear-powered submarines of the Russian Northern Fleet. The project also includes the establishment of a computer-assisted waste monitoring system. In addition, the project involves clearing Sayda Bay of other shipwrecks of the Russian navy. On the German side the project is carried out by the Energiewerke Nord GmbH (EWN) on behalf of the Federal Ministry of Economics and Labour (BMWi). On the Russian side the Kurchatov Institute holds the project management of the long-term interim storage facility in Sayda Bay, whilst the Nerpa Shipyard, which is about 25 km away from the storage facility, is dismantling the submarines and preparing the reactor compartments for long-term interim storage. The technical monitoring of the German part of this project, being implemented by BMWi, is the responsibility of the Federal Institute for Materials Research and Testing (BAM). This paper gives an overview of the German-Russian project and a brief description of solutions for nuclear submarine disposal in other countries. At Nerpa shipyard, being refurbished with logistic and technical support from Germany, the reactor compartments are sealed by welding, provided with biological shielding, subjected to surface treatment and conservation measures. Using floating docks, a tugboat tows the reactor compartments from Nerpa shipyard to the interim storage facility at Sayda Bay where they will be left on the on-shore concrete storage space to allow the radioactivity to decay. For transport of reactor compartments at the shipyard, at the dock and at the storage facility, hydraulic keel blocks, developed and supplied by German subcontractors, are used. In July 2006 the first stage of the reactor compartment storage facility was commissioned and the first seven reactor compartments have been delivered from Nerpa shipyard. Following transports of reactor compartments to the storage facility are expected in 2007.

scholarly journals Measurable Cytokine Concentrations in Pig Seminal Plasma Are Modified by Semen Handling and Storage

Biology ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 276
Author(s):  
Lorena Padilla ◽  
Isabel Barranco ◽  
Inmaculada Parrilla ◽  
Xiomara Lucas ◽  
Heriberto Rodriguez-Martinez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Biological Fluids ◽  
Storage Conditions ◽  
Reliable Measurement ◽  
Short Term ◽  
Gm Csf ◽  
Long Term Storage ◽  
And Storage ◽  
Term Storage

Sample handling and storing are critical steps for the reliable measurement of circulating biomolecules in biological fluids. This study evaluates how cytokine measurements in pig seminal plasma (SP) vary depending on semen handling and SP storage. Thirteen cytokines (GM-CSF, IFNγ, IL-1α, IL-1β, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18 and TNFα) were measured using Luminex xMAP® technology in individual seminal plasma (SP) samples (n = 62) from healthy breeding boars. Three separate experiments explored the delay (2 h and 24 h) in SP collection after ejaculation (Experiment 1) and SP storage, either short-term (5 °C, −20 °C and −80 °C for 72 h, Experiment 2) or long-term (at −20 °C and −80 °C for two months, Experiment 3), before analysis. Levels in fresh SP-samples were used as baseline control values. Delays in SP harvesting of up to 24 h did not substantially impact SP cytokine measurements. Some cytokines showed instability in stored SP samples, mainly in long-term storage. Ideally, cytokines in pig SP should be measured in fresh samples harvested within 24 h after ejaculation. If storage of SP is imperative, storage conditions should be adjusted for each cytokine.

scholarly journals Comparison of methods for pre-processing, exosome isolation, and RNA extraction in unpasteurized bovine and human milk

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257633
Author(s):  
Sanoji Wijenayake ◽  
Shafinaz Eisha ◽  
Zoya Tawhidi ◽  
Michael A. Pitino ◽  
Michael A. Steele ◽  
...  
Keyword(s):  
Human Milk ◽  
Biological Fluid ◽  
Rna Extraction ◽  
Bovine Milk ◽  
Protein Markers ◽  
Long Term Storage ◽  
Transmission Electron ◽  
Cellular Debris ◽  
Term Storage

Milk is a highly complex, heterogeneous biological fluid that contains non-nutritive, bioactive extracellular vesicles called exosomes. Characterization of milk-derived exosomes (MDEs) is challenging due to the lack of standardized methods that are currently being used for milk pre-processing, storage, and exosome isolation. In this study, we tested: 1) three pre-processing methods to remove cream, fat, cellular debris, and casein proteins from bovine milk to determine whether pre-processing of whole milk prior to long-term storage improves MDE isolations, 2) the suitability of two standard exosome isolation methods for MDE fractionation, and 3) four extraction protocols for obtaining high quality RNA from bovine and human MDEs. MDEs were characterized via Transmission Electron Microscopy (TEM), Nanoparticle Tracking Analysis (NTA), and western immunoblotting for CD9, CD63, and Calnexin protein markers. We also present an optimized method of TEM sample preparation for MDEs. Our results indicate that: 1) Removal of cream and fat globules from unpasteurized bovine milk, prior to long-term storage, improves the MDE yield but not purity, 2) Differential ultracentrifugation (DUC) combined with serial filtration is better suited for bovine MDE isolation compared to ExoQuick (EQ) combined with serial filtration, however both methods were comparable for human milk, and 3) TRIzol LS is better suited for RNA extraction from bovine MDEs isolated by EQ and DUC methods. 4) TRIzol LS, TRIzol+RNA Clean and Concentrator, and TRIzol LS+RNA Clean and Concentrator methods can be used for RNA extractions from human MDEs isolated by EQ, yet the TRIzol LS method is better suited for human MDEs isolated by DUC. The QIAzol + miRNeasy Mini Kit produced the lowest RNA yield for bovine and human MDEs.

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