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Conference abstract2013Peer reviewed

Profile of seminal plasma proteins in relation to boars fertility – a pilot study

Madej, Andrzej; Madej, Malgorzata


Semen quality is of great importance, thus markers for reduced fertility have potential economic value. While sperm features are well related to boars’ fertility, the significance of proteins in seminal plasma is not yet fully understood. The objective was to evaluate differences in the FPLC (fast protein liquid chromatography) protein profile of seminal plasma from boars with normal and reduced fertility.

This study included seminal plasma from one ejaculate from each of nine Danish DanBred boars. Six boars displayed normal fertility and three boars displayed reduced fertility. Seminal plasma proteins were separated on column HiPrep 16/10 Heparin FF, by FPLC with some modifications. The column was used directly on ÄKTAdesign™ systems with UNICORN™ software for data calculations on line. Samples of seminal plasma (0.5 ml) were injected through a valve with 0.5 ml sampling loop. Separation of seminal plasma from each boar was repeated four times. The non- heparin binding proteins, the first peak (F1) were eluted with 0.02 M Tris-HCl buffer containing 0.156 M NaCl, pH 7.5. The proteins adsorbed on heparin, the second peak (F2), were eluted using NaCl gradient from 0.156 M to 1.5 M in 0.02 M Tris-HCl buffer, pH 7.5. The used flow rate was 1 ml/min. Yield of eluted proteins was evaluated by measurement of absorbance at 280 nm and 2-ml fractions were collected, lyophilized and stored at -20°C. The following parameters were recorded: total area of peak F1, total area of peak F2, percentage of area for F1, percentage of area for F2, height of peak F1, height of peak F2, total area, ratio between F2 area and F1 area and ratio between height of F1 and F2 (F1/F2 Height). Data was analyzed descriptively using crude means. Profiles from boars with normal fertility served as reference profiles in the comparison to profiles from boars that displayed reduced fertility.

The results indicated differences between boars with high and reduced fertility. One boar with a reduction of 6.5 piglets per litter did not differ in the FPLC profile. Two boars with a reduction of 3 (boar A) and 3.5 (boar B) piglets per litter differed in the FPLC profile compared with boars displaying normal fertility. Results from Boar A showed a reduced height of peak F2 which resulted in the increased ratio of F1/F2 Height. Profile for boar B showed a two fold increase of the height of peak F1 and a corresponding increased ratio of F1/F2 Height.

In conclusion, changes in the height of peaks and ratio between peaks mirror a relation between non-heparin binding (F1 peak) and heparin-binding proteins (F2 peak) in boar seminal plasma. The relation between these two groups of proteins in seminal plasma might affect fertility. A rapid separation of boar seminal plasma on FPLC has potential to be applied as a screening for fertility.

This study was partly funded by Danish DanBred AI-stations, and Ministry of Food, Agriculture and Fisheries of Denmark grant no. 3663-D-10-00461.

Published in

Title: Program and abstract book

Publisher: 9th International Conference on Pig Reproduction


9th International Conference on Pig Reproduction