Seminal plasma addition attenuates the dilution effect in bovine sperm

The substantial decrease of sperm quality after cryopreservation remains an important issue in the artificial insemination industry. Sperm coating with Triladyl® (Minitübe, Tiefenbach, Germany) during ejaculation can preserve sperm characteristics and oocyte penetrating capacity of fresh bovine spermatozoa stored in egg yolk diluent for up to 6 days (De Pauw et al. 2003 Theriogenology 59, 1109–1122). Since collecting semen in a tube containing egg yolk-Tris extender (sperm coating) limits the period of contact between spermatozoa and seminal plasma, the present experiment was conducted to assess if this slightly adjusted method of sperm collection could also have a significant effect on bovine sperm quality after cryopreservation. Semen of five young Holstein Friesian bulls was collected by means of an artificial vagina connected to an empty tube (Group 1; five ejaculates per bull) or a tube containing 4 mL of an egg yolk-Tris extender (Groups 2 and 3; each five ejaculates per bull). The semen samples of Group 1 were conventionally diluted in straws (60 × 106 sperm/mL), frozen, and stored in liquid nitrogen. The samples of Group 3 were centrifuged, and after removing diluent and seminal plasma, the sperm pellet was conventionally diluted and processed. The samples of Group 2 were processed without removal of the supernatant. After thawing each ejaculate was analyzed for average path velocity (VAP), beat cross frequency (BCF), and progressive motility (PROG) using CASA (Minitübe, Tiefenbach, Germany). Furthermore, the membrane integrity of each sample was evaluated using fluorescent SYBR®–14/PI staining (BD Biosciences, Erembodegem, Belgium). All parameters were compared among the three groups of sperm using univariate analysis of variance (SPSS 12.0; SPSS, Inc., Chicago, IL, USA). No significant differences could be observed among the three groups for all of the evaluated sperm characteristics (Table 1). A significant effect of the bull could be determined for all analyzed parameters (P ≤ 0.02), except for the percentage of moribund cells. Nevertheless, the group-bull interaction was never statistically significant. Coating bovine sperm with an egg yolk-Tris extender during ejaculation cannot prevent the substantial deterioration of the spermatozoa that occurs during freezing and thawing since this method of sperm collection does not significantly influence the motility parameters or the membrane integrity after thawing. Table 1. VAP, BCF, PROG, and percentage of membrane-intact, dead, and moribund spermatozoa for the three groups of sperm This research was supported by IWT (no. IWT/020727).

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

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab201 ◽

2012 ◽

Vol 24 (1) ◽

pp. 213 ◽

Cited By ~ 1

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.

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Kinin-induced stimulation of bovine sperm motility and its relation to seminal plasma tissue kallikrein

Immunopharmacology ◽

10.1016/0162-3109(96)00029-x ◽

1996 ◽

Vol 33 (1-3) ◽

pp. 137-139 ◽

Cited By ~ 7

Author(s):

Boris Somlev ◽

Kamel Helili

Keyword(s):

Sperm Motility ◽

Seminal Plasma ◽

Tissue Kallikrein ◽

Bovine Sperm ◽

Stimulation Of

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Is addition or removal of seminal plasma able to compensate for the dilution effect of buffalo semen?

Andrologia ◽

10.1111/and.14123 ◽

2021 ◽

Author(s):

V. Arjun ◽

Pradeep Kumar ◽

Ravi Dutt ◽

Amit Kumar ◽

Renu Bala ◽

...

Keyword(s):

Seminal Plasma ◽

Dilution Effect ◽

Buffalo Semen

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277 EFFECTS OF SEMINAL PLASMA ON SEX-SORTING BOVINE SPERM

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab277 ◽

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.

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The Physiology of Sea-Urchin Spermatozoa Action of Versene

Journal of Experimental Biology ◽

10.1242/jeb.31.2.252 ◽

1954 ◽

Vol 31 (2) ◽

pp. 252-259

Author(s):

LORD ROTHSCHILD ◽

A. TYLER

Keyword(s):

Trace Metals ◽

Sea Urchin ◽

Seminal Plasma ◽

Sea Water ◽

Dilution Effect ◽

Dense Suspension

1. The effect of adding sea water containing different concentrations of versene to suspensions of sea-urchin spermatozoa (Echinus esculentus) has been investigated, as regards their respiration, motility and fertilizing capacity. 2. The respiratory Dilution Effect is progressively reduced and finally abolished when, instead of sea water, sea water containing 10-6, 10-5, 10-4 or 10-3 M versene is added to sperm suspensions. 3. At the same time versene greatly delays the senescence of the spermatozoa, both as regards their motility and fertilizing capacity. For example, after seventeen hours, a 105 sperm/ml. suspension in sea water containing 10-3 M versene has 125 times the fertilizing capacity of a suspension containing 107 sperm/ml. without versene. 4. The change in the ratio of seminal plasma to sea water which occurs when a dense suspension is diluted does not explain the Dilution Effect. 5. These results are discussed in relation to the hypothesis which accounts for the Dilution Effect in terms of trace metals, particularly copper, normally occurring in sea water, and the amounts available per spermatozoon under various conditions of dilution.

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Effects of Dietary Supplementation of Conjugated Linoleic Acids and Their Inclusion in Semen Extenders on Bovine Sperm Quality

Animals ◽

10.3390/ani11020483 ◽

2021 ◽

Vol 11 (2) ◽

pp. 483

Author(s):

Mohammed S. Liman ◽

Vittoria Franco ◽

Claudia L. Cardoso ◽

Valentina Longobardi ◽

Bianca Gasparrini ◽

...

Keyword(s):

Seminal Plasma ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Concentration ◽

Conjugated Linoleic Acids ◽

Factor I ◽

Bovine Sperm ◽

Igf I ◽

Semen Extender ◽

Oxidative Species

Isomers of conjugated linoleic acid (CLA) enhances circulating insulin-like growth factor I (IGF-I) levels. Furthermore, fertility rate of breeding bulls is positively correlated to seminal plasma IGF-I concentration. Our objective was to evaluate the effect of dietary CLA supplementation and inclusion to the semen extender on bovine semen quality and freezability. Fourteen bulls, randomly assigned to control (CTL) and CLA (50 g/day) groups, were supplemented for 10 weeks. Samples were collected at Weeks −2 (before supplementation), 0, 4, 6 (during supplementation), 10, and 11 (after supplementation). Blood and seminal plasma were analyzed for IGF-I; the ejaculates were frozen in the following subgroups: CTL (no addition to semen extender), CLA c9, t11 (50 µM), CLA c9, t11 (100 µM), CLA t10, c12 (50 µM), CLA t10, c12 (100 µM), and CLA mix (50 µM each of CLA c9, t11 and CLA t10, c12). Sperm motility, morphology, viability, mitochondrial membrane potential, and reactive oxidative species were assessed. CLA supplementation decreased ejaculates’ total volume, increased sperm concentration, beat cross frequency, and decreased oxidative stress; it also increased plasma and seminal plasma IGF-I levels compared to the CTL. The inclusion of CLA c9, t11 100 µM and CLA mixture in the extender increased live spermatozoa percentage post-thawing compared to other groups. Our results show a beneficial effect of CLA supplementation on semen quality; however, further studies evaluating fertilization rates are necessary to corroborate the results.

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Seminal plasma addition protects the viability of diluted bovine sperm

Theriogenology ◽

10.1016/s0093-691x(99)91903-9 ◽

1999 ◽

Vol 51 (1) ◽

pp. 344 ◽

Cited By ~ 1

Author(s):

D.L Garner ◽

C.A Thomas ◽

C.E Marshall ◽

J.M DeJarnette ◽

C.H Allen

Keyword(s):

Seminal Plasma ◽

Bovine Sperm

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