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Number of results: 9
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Abstract

The aim of this study was to measure the NO level in boar semen held in a liquid state and to determine its putative relation to spermatozoa motility, plasma membrane integrity, mitochondrial membrane potential and ATP content. Generally, the percentage of spermatozoa which generated nitric oxide gradually increased, while NO level in the surrounding medium declined during the liquid preservation. NO generation in semen preserved in BTS was higher as compared to those in Androhep®Plus. We demonstrated the positive correlation between the NO level in fresh spermatozoa and their quality. We also showed negative correlation between nitric oxide level in spermatozoa preserved in BTS and sperm cells motility as well as plasma membrane integrity. Results obtained in this study confirm that NO may affect sperm physiology in a dualistic manner.
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Authors and Affiliations

A. Orzołek
Ł. Zasiadczyk
P. Wysocki
W. Kordan
P. Krysztofiak
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Abstract

In individual dogs, despite good quality of raw sperm, some parameters are significantly changed after thawing, which cannot be predicted. We therefore investigated whether motility parameters objectively obtained by CASA, membrane integrity (MI), cell morphology or a combination are suitable to improve the prediction of bad post-thaw quality. For this purpose 250 sperm analysis protocols from 141 healthy stud dogs, all patients introduced for sperm cryopreservation, were evaluated and a Classification and Regression Tree (CART) -analysis performed. The sperm was routinely collected, analysed, and frozen by using a modified Uppsala system. After thawing, data were routinely examined by using CASA, fluorescent microscopy for membrane integrity (MI) and Hancock’s fixation for evaluation of cell morphology. Samples were sorted by post-thaw progressive motility (P) in good (P > / = 50%, n=135) and bad freezers (P<50%, n=115). Among bad freezers, 73.9% showed in addition post-thaw total morphological abberations of >40% and/or MI <50%.

Bad freezers were significantly older than good freezers (p<0.05). Progressive motility (P), velocity curvilinear (VCL), mean coefficient (STR), and linear coefficient (LIN) were potential predictors for post-thaw sperm quality since specifity was best (85.8%) and sensitivity (75.4 %) and accuracy (80.4 %) good. For these objectively measured raw sperm parameters, cut-off values were calculated allowing prediction of bad post-thaw results with high accuracy: P = 83.1 % VCL = 161.3 µm/sec, STR = 0.83 %, and LIN = 0.48 %. Raw sperm samples with values below these cut off values will have below average post-thaw quality with a probability of 85.8%. We conclude that VCL, P, STR and LIN are potential predictors of the outcome of sperm cryopreservation, when combined.

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Authors and Affiliations

S. Schäfer-Somi
A. Tichy
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Abstract

Aim: The aim of this study was to analyze the effect of bovine follicular fluid on the survival, morphology and kinetic parameters of bovine thawed spermatozoa under laboratory conditions.

Materials and methods: The semen from 5 bulls of proven fertility was incubated in follicular and physiological fluid for 8 hours. During this time assessment using the CASA system was performed. At the beginning and the end of incubation process evaluation by flow cytometry was conducted.

Results: The results of the sperm motility assessment showed a significant decrease in the analyzed parameters both in the follicular and physiological fluid. A significant reduction in all parameters characterizing movement properties in the semen incubated in the follicular fluid was found. In the physiological fluid, a similar trend was demonstrated only for the following proper- ties: VAP, VSL, VCL, ALH, BCF. A significant difference was found for both fluids in: VCL (p=0.026), ALH (p=0.038) and LIN (p<0.001) at the beginning of incubation. The results of the plasma membrane integrity assessment showed a statistically significant increase in the percent- age of dying sperm at the 8th hour of the incubation in the follicular fluid. In the case of semen incubation in physiological fluid, a statistically significant decrease in the percentage of live non-damaged cells was found with a simultaneous increase in the subpopulation of undamaged dead cells.

Conclusions: Follicular fluid rapidly accelerates the capacitation process. The results of flow cytometry support the hypothesis concerning the ability of follicular fluid to prolong sperm sur- vival.

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Authors and Affiliations

J. Mrowiec
J. Twardoń
A. Bartoszewicz
W. Niżański
M. Ochota
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Abstract

This is the first study to identify 23 protein spots corresponding to 13 proteins in mitochondria isolated from rabbit epididymal spermatozoa. In the group of protein spots identified in stress-induced samples, the abundance of 20 protein spots increased, whereas the abundance of three protein spots (GSTM3, CUNH9orf172, ODF1) decreased relative to the control. The results of this study provide valuable inputs for future research into the molecular mechanisms implicated in pathological processes during oxidative stress (OS).
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Authors and Affiliations

M. Lecewicz
1
W. Kordan
1
K. Górski
2
S. Kondracki
2

  1. Department of Animal Biochemistry and Biotechnology, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-719 Olsztyn, Poland
  2. Institute of Animal Production and Fisheries, Siedlce University of Natural Sciences and Humanities, Prusa 14, 08-110 Siedlce, Poland
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Abstract

The aim of this study was to compare the cryoprotective effects of egg yolk from different avian species (hen, goose and quail) on post-thaw quality of dog semen. Total motility (TMOT) and progressive motility (PMOT) of frozen-thawed spermatozoa were not significantly differed among the extenders, but were higher in the quail-egg yolk based extender compared with extender containing hen or goose egg yolk. It was found that post-thaw sperm motion parameters, velocity VCL and ALH, were significantly higher in the quail-egg yolk based extender. No marked differences in post-thaw sperm plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP) were observed among the extenders. In conclusion, the results of the present study suggest that goose or quail egg yolk is a suitable alternative to hen egg yolk for the cryopreservation of dog semen.
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Bibliography

Bathgate R, Maxwell WMC, Evans G (2006) Studies on the effect of supplementing boar semen cryopreservation media with different avian egg yolk types on in vitro post-thaw sperm quality. Reprod Domest Anim 41: 68-73.
Kulaksiz R, Cebi C, Akcay E, Daskin A (2010) The protective effect of egg yolk from different avian species during the cryopreservation of Karayaka ram semen. Small Rumin Res 88: 12-15.
Parks JE, Lynch DV (1992) Lipid composition and thermotropic phase behaviour of boar, bull, stallion, and rooster sperm membranes. Cryobiology 29: 255-266.
Rijsselaere T, Van Soom A, Maes D, Niżański W (2012) Computer-assisted sperm analysis in dogs and cats: an update after 20 years. Reprod Domest Anim 47 (Suppl 6): 204-207.
Santiago-Moreno J, Coloma MA, Toledano-Díaz A, Gómez-Brunet A, Pulido-Pastor A, Zamora-Soria A, Carrizosa JA, Urrutia B, López-Sebastián A (2008) A comparison of the protective action of chicken and quail egg yolk in the cryopreservation of Spanish ibex epididymal spermatozoa. Cryobiology 57: 25-29.
Strzeżek R, Polakiewicz P, Kordan W (2015) The effect of two packaging systems on the post-thaw characteristics of canine sperm. Pol J Vet Sci 18: 249-253.
Su L, Li X, Quan J, Yang S, Li Y, He X, Tang X (2008) A comparison of the protective action of added egg yolks from five avian species to the cryopreservation of bull sperm. Anim Reprod Sci 104: 212-219.
Trimeche A, Anton M, Renard P, Gandemer G, Tainturier D (1997). Quail egg yolk: a novel cryoprotectant for the freeze preservation of Poitou jackass sperm. Cryobiology 34: 385-393.
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Authors and Affiliations

R. Strzeżek
1
A. Reksa
1

  1. Department of Animal Biochemistry and Biotechnology, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-718 Olsztyn-Kortowo, Poland
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Abstract

The purpose of this study is to determine the effects of proanthocyanidin (PA) on spermatological parameters and testicular toxicity in male rats exposed to glyphosate (GLP). In our study, four groups were formed out of 24 male rats, each group would include 6 rats. The rats in the PA group were given a dose of 400 mg/kg/day dissolved in DMSO via gastric gavage. The rats in the GLP+PA groups were first given GLP at the LD50/10 dose of 787.85 mg/kg/day, followed by administering PA at a dose of 400 mg/kg/day dissolved in DMSO via gastric gavage. The rats in the GLP group were given GLP at the LD50/10 dose of 787.85 mg/kg/day dissolved in DMSO via gastric gavage. It was determined that in terms of motility, in comparison to the control group, the decreases in the GLP group and the increases in the PA and GLP+PA groups were statistically significant (p<0.001). The administration of GLP increased DNA damage compared to the control group, but the GLP+PA and PA applications reduced DNA damage (p<0.001). The analysis of testosterone levels indicated a statistically significant reduction in the GLP group compared to the other groups. Consequently, it was determined that PA effectively prevented the decreases in the spermatological parameters lowered as a result of GLP exposure and the oxidative stress and toxicity in testicular tissue.
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Bibliography

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Authors and Affiliations

F. Avdatek
1
M. Kirikkulak
1
D. Yeni
1

  1. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, University of Afyon Kocatepe, 03030, Afyonkarahisar, Turkey
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Abstract

In this study the quality of total RNA, isolated from fresh spermatozoa, was compared between boars with good and poor semen freezability (GSF and PSF, respectively). Semen from 3 boars with GSF exhibited significantly higher total motility, mitochondrial function, plasma membrane integrity and reduced lipid peroxidation compared with 3 boars with PSF after cryo- preservation. There were variations in the quality of RNA isolated from spermatozoa of boars with GSF and PSF. Boars with GSF exhibited mainly full-length, intact RNA, whereas substantial amounts of degraded RNA were detected in spermatozoa from boars with PSF. Further under- standing of the biological relevance of RNAs in sperm function is critical to improve the freezabil- ity of boar semen.

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Authors and Affiliations

L. Fraser
P. Brym
M. Mogielnicka-Brzozowska
K. Wasilewska

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