Morphometry of Boar Spermatozoa in Semen Stored at 17 °C-The Influence of the Staining Technique

Abstract

The aim of the study was to assess the morphometry of sperm during storage of liquid boar semen at 17 °C. An attempt was also made to evaluate the suitability of three staining methods for assessment of boar sperm morphometry. The study was carried out on 20 Landrace boars. Semen was collected from the boars every 5 days by the manual method. Four ejaculates from each boar were analysed (80 ejaculates in total). Analyses were performed five times: at 1 h, 24 h, 48 h, 96 h, and 168 h after semen collection. Blisters with insemination doses were opened immediately before the analyses. From each insemination dose, smears were prepared for morphometric evaluation of sperm, which were stained by three methods (eosin-nigrosin-EN, eosin-gentian-EG, and SpermBlue-SB). Morphometric measurements of 15 randomly selected sperm with normal morphology were performed on each slide. The morphometric measurements included the following parameters: sperm head length, width, area, and perimeter; tail length; and total sperm length. The results of the morphometric measurements were used to calculate the head shape index. The morphometric dimensions of the sperm were shown to change during storage of semen at 17 °C. The extent of these changes, however, depended on the staining method used, as the three methods result in different morphometric dimensions of sperm, in the case of both the head and the tail. In the slides stained by the eosin-nigrosin method, the dimensions of the head and tail were smaller at every time of storage than in the slides stained by the SpermBlue and eosin-gentian methods.

Keywords: boar; semen storage time; sperm morphometry.

Figure 1

Morphometric dimensions of the heads of sperm stained by the eosin-nigrosin (EN), eosin-gentian (EG), and SpermBlue (SB) methods depending on semen storage time ((A)—length, (B)—width, (C)—area, (D)—perimeter). Bars with different letters mean statistically significant values (p ≤ 0.05).

Figure 2

Length of sperm stained by the eosin-nigrosin (EN), eosin-gentian (EG), and SpermBlue (SB) methods depending on semen storage time ((A)—tail length, (B)—total sperm length). Bars with different letters mean statistically significant values (p ≤ 0.05).