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 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.

277 EFFECTS OF SEMINAL PLASMA ON SEX-SORTING BOVINE SPERM

2011 ◽  
Vol 23 (1) ◽  
pp. 236
Author(s):  
C. A. Burroughs ◽  
R. W. Lenz ◽  
K. M. Evans ◽  
J. K. Graham ◽  
G. E. Seidel
Keyword(s):  
Seminal Plasma ◽  
Sperm Concentration ◽  
Hoechst 33342 ◽  
Future Studies ◽  
Food Dye ◽  
Bovine Sperm ◽  
Initial Ph ◽  
Sperm Membrane ◽  
Red Dye ◽  
Artificial Vagina

This experiment evaluated how characteristics of bovine ejaculates affect the sortability of X- and Y-bearing spermatozoa. Ejaculates were collected by artificial vagina, 2 each from 10 bulls with an average of 1 h between collections. Only ejaculates with at least 60% motile and 70% normal sperm were used. Semen was centrifuged at 1 000 × g for 15 min to separate sperm from seminal plasma; seminal plasma was clarified by 10 min of additional centrifugation at 2 000 × g. Sperm were rediluted to 160 million sperm/mL with TALP (pH 7.4) and 0, 5, 10, or 20% seminal plasma, from the same ejaculate or reciprocally from first/second ejaculates. Following incubation with Hoechst 33342 for 45 min, an equal volume of TALP (pH 5.5) containing red food dye was added, and sperm were analysed on a MoFlo (Dako, Glostrup, Denmark) flow cytometer for percentage live-oriented cells, X sort rate, coincidence rate, percentage dead or dying (sperm membrane permeable to red dye), and splitability (peaks to valley ratio). The percentage live-oriented sperm was higher for treatments with 0% seminal plasma (64.4%) than for 5 (59.6%), 10 (59.0%), and 20% (57.8%) seminal plasma (P < 0.01). The percentage live-oriented sperm was higher for second (63.0%) than for first ejaculates (56.2%). Sort rate was higher for 0% seminal plasma and second ejaculates (P < 0.05). Dead/dying rates were lower for 0% (16.5%) than for 5 (21.9%), 10 (23.6%), or 20% (23.4%) seminal plasma (P < 0.003); and for first ejaculates (25.9%) compared with second ejaculates (18.2%). Seminal plasma percentage and ejaculate had no effect on splitability, and there was no difference in sorting parameters whether the seminal plasma was from the first or second ejaculate. The initial sperm concentration of the ejaculate (range, 1.3 to 3.4 billion sperm/mL) did not affect any sperm sorting parameter. Thus, effects of sperm concentration on sortability need to be reconsidered, as current practice is to select ejaculates to sort based on initial sperm concentration. The initial pH, percentage morphological abnormalities, initial seminal plasma pH, and age of bull did not affect sorting parameters. In conclusion, the presence of seminal plasma during staining and sorting may decrease sort rate and percentage of live-oriented cells, as well as increase death rate. In addition, sorting second ejaculates may be more advantageous than sorting first ejaculates. Future studies are needed to determine if the results hold true if sperm are stored for several hours and how the various factors affect sperm post-thaw viability.

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 Metabolite Profiling of Pig Seminal Plasma Identifies Potential Biomarkers for Sperm Resilience to Liquid Preservation

2021 ◽  
Vol 9 ◽  
Author(s):  
Yentel Mateo-Otero ◽  
Pol Fernández-López ◽  
Jordi Ribas-Maynou ◽  
Jordi Roca ◽  
Jordi Miró ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Liquid Storage ◽  
Dependent Behavior ◽  
Downstream Gene Expression ◽  
Cell Signaling Pathways ◽  
C Nmr

Metabolomic approaches allow the study of downstream gene expression events since metabolites are considered as the products of cell signaling pathways. For this reason, many studies in humans have already been conducted to determine the influence of the metabolites present in seminal plasma (SP) on sperm physiology, and to identify putative biomarkers. However, in livestock species, these relationships are yet to be uncovered. Thus, the present study aimed to explore: (i) if concentrations of metabolites in pig SP are related to sperm quality and functionality, and (ii) if they could predict the sperm resilience to liquid storage at 17°C. To this end, 28 ejaculates were individually collected and split into three aliquots: one was used for SP analysis through nuclear magnetic resonance (NMR) spectroscopy; another served for the evaluation of sperm concentration and morphology; and the last one was utilized to determine sperm functionality parameters using computer-assisted sperm analysis (CASA) and flow cytometry after 0 h and 72 h of liquid-storage at 17°C. NMR analysis allowed the identification and quantification of 23 metabolites present in pig SP which, except for fumarate, were not observed to follow a breed-dependent behavior. Moreover, specific relationships between metabolites and sperm variables were identified: (i) glutamate, methanol, trimethylamine N-oxide, carnitine, and isoleucine were seen to be related to some sperm quality and functionality parameters evaluated immediately after semen collection; (ii) leucine, hypotaurine, carnitine and isoleucine were found to be associated to the sperm ability to withstand liquid storage; and (iii) Bayesian multiple regression models allowed the identification of metabolite patterns for specific sperm parameters at both 0 h and 72 h. The identification of these relationships opens up the possibility of further investigating these metabolites as potential sperm functional biomarkers.

The effect of different extenders on the sperm motilityand viability of frozen Turkey semen

2016 ◽  
Author(s):  
Mehmet K. Kuzlu ◽  
Atilla Taskin
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Concentration ◽  
Dimethyl Sulphoxide ◽  
Freezing And Thawing ◽  
Long Term Storage ◽  
Nitrogen Vapor ◽  
Storage Methods ◽  
Term Storage

This study was conducted to develop long-term storage methods for turkey semen using different extenders. During the study, the massage method was used twice a week to collect the semen from five turkeys, a total of 44 times. The collected fresh semen’s average ejaculate quantity, sperm concentration, motility and vitality values were determined as 0.22±0.01 ml, 3.5±0.17 x109 sp/ml, 77.0±1.44% and 86.2±0.95 % respectively. In this study, glucose (G) including 5% Dimethyl sulphoxide (DMSO) cryoprotective, tris-glucose (TG), lactated Ringer’s (LR), and lactated Ringer’s glucose (LRG) extenders were used. The Turkey semen was combined and divided into four equal parts, and subsequently diluted at a ratio of 1:3 and equilibrated at +4°C for 90 minutes. Following equilibration, the samples were frozen in liquid nitrogen vapor to - 80°C for five minutes, and stored at -196°C in liquid nitrogen. After freezing and thawing, the highest motility value was obtained from the G extender (43.3% ±1.62%) followed by LRG (24.6±1.53%), LR (12.6±0.92%) and TG (12.2±0.66%). The vitality values were recorded as 55.8±1.89%, 23.8±1.58%, 21.5±1.10% and 36.5±1.59% respectively. The motility and vitality values were significantly (p less than 0.01) more for the glucose extender than those for other extenders. Therefore, it was concluded that glucose extender is better option for the long-term storage of Turkey semen.

scholarly journals Effect of different extenders on ram sperm traits during storage

2012 ◽  
Vol 60 (6) ◽  
pp. 111-116 ◽  
Author(s):  
Zdeňka Hegedűšová ◽  
Ladislav Štolc ◽  
František Louda ◽  
Libor Čunát ◽  
Jan Vejnar
Keyword(s):  
Sperm Motility ◽  
Sperm Concentration ◽  
Short Term ◽  
Long Term Storage ◽  
Semen Volume ◽  
Reproduction Season ◽  
Ram Sperm ◽  
Sperm Traits ◽  
Term Storage

The aim of the study was to test commercial extenders used for short-term and long-term sperm preservation. Semen was collected in the reproduction season, i.e. from June to December. The ejaculates were obtained from single services and the routine analysis of the semen was performed immediately after the collection. The examination included semen volume, colour and texture, sperm concentration and motility, ejaculate turbulence and percentage of sperm with abnormal morphology. The semen was diluted with an extender in the ratio of 1:4. The processed semen was transported in an insulated container at 16–18 °C to the laboratory and stored in a stationary thermostat under the same temperature. Sperm motility tests were performed 24, 48, 72 and 96 hours after the placement in to thermostat. Ejaculates diluted with Ovipro, Optidyl, Triladyl and Andromed CSS gave very good results of viability (81.23 %–83.41 %) after 24 hours of storage. After 48 hours, Ovipro, Andromed, Optidyl and Triladyl gave values above 75 %. The Triladyl extender proved to be a good stabilizing agent, showing consistent results during a long-term storage. It was chosen as a control one for overall assessment. Other preservation media did not show any improving or worsening effects. The extender Ovipro showed a high motility effect in the first 48 hours only, and hence it appears to be the best solution for the short-term preservation.

Modelling of Moisture Movement in Wood during Outdoor Storage

2001 ◽  
Vol 6 (2) ◽  
pp. 3-14 ◽  
Author(s):  
R. Baronas ◽  
F. Ivanauskas ◽  
I. Juodeikienė ◽  
A. Kajalavičius
Keyword(s):  
Experimental Data ◽  
Finite Difference ◽  
Climatic Conditions ◽  
Two Dimensional ◽  
Moisture Movement ◽  
Finite Difference Technique ◽  
Long Term Storage ◽  
Space Formulation ◽  
Term Storage

A model of moisture movement in wood is presented in this paper in a two-dimensional-in-space formulation. The finite-difference technique has been used in order to obtain the solution of the problem. The model was applied to predict the moisture content in sawn boards from pine during long term storage under outdoor climatic conditions. The satisfactory agreement between the numerical solution and experimental data was obtained.

Trehalose: a cryoprotectant that enhances recovery and preserves function of human pancreatic islets after long-term storage

Diabetes ◽  
1997 ◽  
Vol 46 (3) ◽  
pp. 519-523 ◽  
Author(s):  
G. M. Beattie ◽  
J. H. Crowe ◽  
A. D. Lopez ◽  
V. Cirulli ◽  
C. Ricordi ◽  
...  
Keyword(s):  
Pancreatic Islets ◽  
Human Pancreatic Islets ◽  
Long Term Storage ◽  
Term Storage
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